44 results on '"Eyal, Nir"'
Search Results
2. Self-Assembly of DNA Origami Heterodimers in High Yields and Analysis of the Involved Mechanisms
- Author
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Yaron Berger, Eyal Nir, Dinesh Chandra Khara, Miran Liber, Jürgen Jopp, Mary Popov, and Breveruos Sheheade
- Subjects
02 engineering and technology ,Polyethylene glycol ,010402 general chemistry ,01 natural sciences ,Dissociation (chemistry) ,Polyethylene Glycols ,Biomaterials ,chemistry.chemical_compound ,Sticky and blunt ends ,DNA nanotechnology ,DNA origami ,Nanotechnology ,General Materials Science ,Electrophoresis, Agar Gel ,General Chemistry ,DNA ,021001 nanoscience & nanotechnology ,Combinatorial chemistry ,0104 chemical sciences ,Monomer ,chemistry ,Agarose gel electrophoresis ,Nucleic Acid Conformation ,Self-assembly ,0210 nano-technology ,Biotechnology - Abstract
Efficient fabrication of structurally and functionally diverse nanomolecular devices and machines by organizing separately prepared DNA origami building blocks into a larger structure is limited by origami attachment yields. A general method that enables attachment of origami building blocks using 'sticky ends' at very high yields is demonstrated. Two different rectangular origami monomers are purified using agarose gel electrophoresis conducted in solute containing 100 × 10-3 m NaCl, a treatment that facilitates the dissociation of most of the incorrectly hybridized origami structures that form through blunt-end interactions during the thermal annealing process and removes these structures as well as excess strands that otherwise interfere with the desired heterodimerization reaction. Heterodimerization yields of gel-purified monomers are between 98.6% and 99.6%, considerably higher than that of monomers purified using the polyethylene glycol (PEG) method (88.7-96.7%). Depending on the number of PEG purification rounds, these results correspond to about 4- to 25-fold reduction in the number of incorrect structures observed by atomic force microscopy. Furthermore, the analyses of the incorrect structures observed before and after the heterodimerization reactions and comparison of the purification methods provide valuable information on the reaction mechanisms that interfere with heterodimerization.
- Published
- 2019
3. Study of DNA Origami Dimerization and Dimer Dissociation Dynamics and of the Factors that Limit Dimerization
- Author
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Miran Liber, Eyal Nir, Mary Popov, Roman Tsukanov, Yaron Berger, Toma E. Tomov, and Dinesh Chandra Khara
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Materials science ,Dimer ,Kinetics ,02 engineering and technology ,010402 general chemistry ,01 natural sciences ,Dissociation (chemistry) ,Fluorescence ,Biomaterials ,chemistry.chemical_compound ,DNA nanotechnology ,DNA origami ,General Materials Science ,Ions ,General Chemistry ,DNA ,021001 nanoscience & nanotechnology ,Single-molecule experiment ,Molecular machine ,0104 chemical sciences ,Monomer ,chemistry ,Chemical physics ,Thermodynamics ,0210 nano-technology ,Dimerization ,Biotechnology - Abstract
Organizing DNA origami building blocks into higher order structures is essential for fabrication of large structurally and functionally diverse devices and molecular machines. Unfortunately, the yields of origami building block attachment reactions are typically not sufficient to allow programed assembly of DNA devices made from more than a few origami building blocks. To investigate possible reasons for these low yields, a detailed single-molecule fluorescence study of the dynamics of rectangular origami dimerization and origami dimer dissociation reactions is conducted. Reactions kinetics and yields are investigated at different origami and ion concentrations, for different ion types, for different lengths of bridging strands, and for the "sticky end" and "weaving welding" attachment techniques. Dimerization yields are never higher than 86%, which is typical for such systems. Analysis of the dynamic data shows that the low yield cannot be explained by thermodynamic instability or structural imperfections of the origami constructs. Atomic force microscopy and gel electrophoresis evidence reveal self-dimerization of the origami monomers, likely via blunt-end interactions made possible by the presence of bridging strands. It is suggested that this mechanism is the major factor that inhibits correct dimerization and means to overcome it are discussed.
- Published
- 2018
4. DNA Bipedal Motor Achieves a Large Number of Steps Due to Operation Using Microfluidics-Based Interface
- Author
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Eyal Nir, Miran Liber, Yair Glick, Dorit Avrahami, Yaron Berger, Roman Tsukanov, Doron Gerber, and Toma E. Tomov
- Subjects
Computer science ,Surface Properties ,Interface (computing) ,Microfluidics ,Immobilized Nucleic Acids ,General Physics and Astronomy ,Nanotechnology ,02 engineering and technology ,010402 general chemistry ,01 natural sciences ,chemistry.chemical_compound ,Lab-On-A-Chip Devices ,DNA nanotechnology ,Fluorescence Resonance Energy Transfer ,General Materials Science ,business.industry ,General Engineering ,Processivity ,DNA ,Microfluidic Analytical Techniques ,021001 nanoscience & nanotechnology ,Molecular machine ,Single Molecule Imaging ,0104 chemical sciences ,Biomechanical Phenomena ,Nanostructures ,Kinetics ,chemistry ,User interface ,0210 nano-technology ,business ,Computer hardware - Abstract
Realization of bioinspired molecular machines that can perform many and diverse operations in response to external chemical commands is a major goal in nanotechnology, but current molecular machines respond to only a few sequential commands. Lack of effective methods for introduction and removal of command compounds and low efficiencies of the reactions involved are major reasons for the limited performance. We introduce here a user interface based on a microfluidics device and single-molecule fluorescence spectroscopy that allows efficient introduction and removal of chemical commands and enables detailed study of the reaction mechanisms involved in the operation of synthetic molecular machines. The microfluidics provided 64 consecutive DNA strand commands to a DNA-based motor system immobilized inside the microfluidics, driving a bipedal walker to perform 32 steps on a DNA origami track. The microfluidics enabled removal of redundant strands, resulting in a 6-fold increase in processivity relative to an identical motor operated without strand removal and significantly more operations than previously reported for user-controlled DNA nanomachines. In the motor operated without strand removal, redundant strands interfere with motor operation and reduce its performance. The microfluidics also enabled computer control of motor direction and speed. Furthermore, analysis of the reaction kinetics and motor performance in the absence of redundant strands, made possible by the microfluidics, enabled accurate modeling of the walker processivity. This enabled identification of dynamic boundaries and provided an explanation, based on the "trap state" mechanism, for why the motor did not perform an even larger number of steps. This understanding is very important for the development of future motors with significantly improved performance. Our universal interface enables two-way communication between user and molecular machine and, relying on concepts similar to that of solid-phase synthesis, removes limitations on the number of external stimuli. This interface, therefore, is an important step toward realization of reliable, processive, reproducible, and useful externally controlled DNA nanomachines.
- Published
- 2017
5. Developing DNA Nanotechnology Using Single-Molecule Fluorescence
- Author
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Eyal Nir, Roman Tsukanov, Miran Liber, Yaron Berger, and Toma E. Tomov
- Subjects
chemistry.chemical_classification ,DNA, Cruciform ,Computer science ,Lasers ,Biomolecule ,Immobilized Nucleic Acids ,Nanotechnology ,Context (language use) ,DNA ,General Medicine ,General Chemistry ,Single-molecule experiment ,Fluorescence ,Diffusion ,Kinetics ,Förster resonance energy transfer ,chemistry ,DNA nanotechnology ,Fluorescence Resonance Energy Transfer ,Holliday junction ,Nucleic Acid Conformation ,DNA origami ,A-DNA - Abstract
CONSPECTUS: An important effort in the DNA nanotechnology field is focused on the rational design and manufacture of molecular structures and dynamic devices made of DNA. As is the case for other technologies that deal with manipulation of matter, rational development requires high quality and informative feedback on the building blocks and final products. For DNA nanotechnology such feedback is typically provided by gel electrophoresis, atomic force microscopy (AFM), and transmission electron microscopy (TEM). These analytical tools provide excellent structural information; however, usually they do not provide high-resolution dynamic information. For the development of DNA-made dynamic devices such as machines, motors, robots, and computers this constitutes a major problem. Bulk-fluorescence techniques are capable of providing dynamic information, but because only ensemble averaged information is obtained, the technique may not adequately describe the dynamics in the context of complex DNA devices. The single-molecule fluorescence (SMF) technique offers a unique combination of capabilities that make it an excellent tool for guiding the development of DNA-made devices. The technique has been increasingly used in DNA nanotechnology, especially for the analysis of structure, dynamics, integrity, and operation of DNA-made devices; however, its capabilities are not yet sufficiently familiar to the community. The purpose of this Account is to demonstrate how different SMF tools can be utilized for the development of DNA devices and for structural dynamic investigation of biomolecules in general and DNA molecules in particular. Single-molecule diffusion-based Förster resonance energy transfer and alternating laser excitation (sm-FRET/ALEX) and immobilization-based total internal reflection fluorescence (TIRF) techniques are briefly described and demonstrated. To illustrate the many applications of SMF to DNA nanotechnology, examples of SMF studies of DNA hairpins and Holliday junctions and of the interactions of DNA strands with DNA origami and origami-related devices such as a DNA bipedal motor are provided. These examples demonstrate how SMF can be utilized for measurement of distances and conformational distributions and equilibrium and nonequilibrium kinetics, to monitor structural integrity and operation of DNA devices, and for isolation and investigation of minor subpopulations including malfunctioning and nonreactive devices. Utilization of a flow-cell to achieve measurements of dynamics with increased time resolution and for convenient and efficient operation of DNA devices is discussed briefly. We conclude by summarizing the various benefits provided by SMF for the development of DNA nanotechnology and suggest that the method can significantly assist in the design and manufacture and evaluation of operation of DNA devices.
- Published
- 2014
6. Photon-by-Photon Hidden Markov Model Analysis for Microsecond Single-Molecule FRET Kinetics
- Author
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Rashid Khamis, Yaron Berger, Roman Tsukanov, Gilad Haran, Menahem Pirchi, Hadas Volkov, Toma E. Tomov, Dinesh Chandra Khara, and Eyal Nir
- Subjects
0301 basic medicine ,Quantitative Biology::Biomolecules ,Photon ,Scale (ratio) ,Chemistry ,Analytical chemistry ,Estimator ,Function (mathematics) ,Single-molecule FRET ,010402 general chemistry ,01 natural sciences ,0104 chemical sciences ,Surfaces, Coatings and Films ,03 medical and health sciences ,Microsecond ,030104 developmental biology ,Förster resonance energy transfer ,Materials Chemistry ,Physical and Theoretical Chemistry ,Hidden Markov model ,Biological system - Abstract
The function of biological macromolecules involves large-scale conformational dynamics spanning multiple time scales, from microseconds to seconds. Such conformational motions, which may involve whole domains or subunits of a protein, play a key role in allosteric regulation. There is an urgent need for experimental methods to probe the fastest of these motions. Single-molecule fluorescence experiments can in principle be used for observing such dynamics, but there is a lack of analysis methods that can extract the maximum amount of information from the data, down to the microsecond time scale. To address this issue, we introduce H2MM, a maximum likelihood estimation algorithm for photon-by-photon analysis of single-molecule fluorescence resonance energy transfer (FRET) experiments. H2MM is based on analytical estimators for model parameters, derived using the Baum–Welch algorithm. An efficient and effective method for the calculation of these estimators is introduced. H2MM is shown to accurately retrieve...
- Published
- 2016
7. Detailed Study of DNA Hairpin Dynamics Using Single-Molecule Fluorescence Assisted by DNA Origami
- Author
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Eyal Nir, Toma E. Tomov, Miran Liber, Hagai Drory, Roman Tsukanov, Noa Plavner, and Rula Masoud
- Subjects
Chemistry ,Inverted Repeat Sequences ,Immobilized Nucleic Acids ,Fluorescence correlation spectroscopy ,DNA ,Sodium Chloride ,Single-molecule experiment ,Fluorescence ,Surfaces, Coatings and Films ,Gibbs free energy ,Diffusion ,Crystallography ,symbols.namesake ,Förster resonance energy transfer ,Ionic strength ,Metastability ,Fluorescence Resonance Energy Transfer ,Materials Chemistry ,symbols ,Thermodynamics ,DNA origami ,Physical and Theoretical Chemistry - Abstract
The dynamics of two DNA hairpins (5'-TCGCCT-A31-AGGCGA-3' and 5'-TCGCCG-A31-CGGCGA-3') were studied using immobilization-based and diffusion-based single-molecule fluorescence techniques. The techniques enabled separated and detailed investigation of the states and of the transition reactions. Only two states, open and closed, were identified from analysis of the FRET histograms; metastable states with lifetimes longer than the technique resolution (0.3 ms) were not observed. The opening and closing reaction rates were determined directly from the FRET time trajectories, and the Gibbs free energies of these states and of the transition state were calculated using the Kramer theory. The rates, which are undoubtedly of transitions between the fully closed and the fully open states and ranged from 2 to 90 s(-1), were lower (∼10-fold) than the rates previously determined from fluorescence correlation spectroscopy. The heights of the barriers for closing were almost identical for the two hairpins. The barrier for opening the hairpin with the stronger stem was higher (4.3 kJ/mol) than that for the hairpin with the weaker stem, in very good agreement with the difference in stability calculated by the nearest-neighbor method. The barrier for closing the hairpin decreased (∼8 kJ/mol) and the barrier for opening increased (∼4 kJ/mol) with increasing NaCl concentration (10-100 mM), indicating that higher ionic strength stabilizes the folded state with respect to the transition state and stabilizes the transition state relative to the unfolded state. The very good agreements in the dynamics measured for free hairpins, for hairpins anchored to origami, and for hairpins anchored to the coverslip and the very good agreement between the two single-molecule techniques demonstrate that neither the origami nor the coverslip influence the hairpin dynamics, supporting a previous demonstration that origami can serve as a platform for biophysical investigations.
- Published
- 2013
8. Grounding Public Reasons in Rationality: The Conditionally-Compassionate Medical Student and Other Challenges
- Author
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Eyal Nir
- Subjects
Sociology and Political Science ,Rationality ,Sociology ,Positive economics ,Law ,Reciprocity (evolution) ,Social psychology ,Public reason - Abstract
Gillian Hadfield and Stephen Macedo argue that late-Rawlsian stability for the right reasons, that is, stability based on participants’ reciprocal cooperation, can arise even if participants start out only economically rational and indifferent to justice. As they explain, even purely rational actors have an interest in having a neutral “shared logic” to coordinate decentralized enforcement of social cooperation and in internalizing that logic. Once developed and internalized, they add, that logic renders their reasoning public, and their persons, reasonable and responsive to Rawlsian justice considerations. Hadfield and Macedo’s account seeks to lend Rawlsian contractualism a contractarian foundation. I raise five challenges to their account, focusing on the sense in which actors “start out” purely rational; on illiberal means that may serve rational citizens’ joint ends; on the counterexample of Martin Luther King; on the difficulty of specifying the mechanism for shared logic-internalization; and on the further counterexample of a medical student who internalizes compassion, only upon the condition that it serves his rational aims.
- Published
- 2012
9. Studying the Structural Dynamics of Bipedal DNA Motors with Single-Molecule Fluorescence Spectroscopy
- Author
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Miran Liber, Eyal Nir, Rula Masoud, Noa Plavner, Roman Tsukanov, and Toma E. Tomov
- Subjects
Work (thermodynamics) ,Chemistry ,Lasers ,General Engineering ,Phase (waves) ,General Physics and Astronomy ,Nanotechnology ,DNA ,Single-molecule experiment ,Kinetics ,Motion ,chemistry.chemical_compound ,Spectrometry, Fluorescence ,Förster resonance energy transfer ,Yield (chemistry) ,DNA nanotechnology ,Fluorescence Resonance Energy Transfer ,General Materials Science ,Spectroscopy ,Biological system ,Fluorescent Dyes - Abstract
We present a test case example of a detailed single-molecule fluorescence study of one of the most sophisticated and complex DNA devices introduced to date, a recently published autonomous bipedal DNA motor. We used the diffusion-based single-molecule Förster resonance energy transfer technique, coupled to alternating laser excitation (sm-FRET-ALEX), to monitor the motor assembly and operation. The study included verification of the formation of the correct structures, and of the correct motor operation, determination of the formation and stepping reaction yields, and identification of side products. Finally, the mechanisms of the motor assembly and operation were elucidated by measuring the reaction kinetics profile of track-walker binding and of lifting of the walker's leg upon fuel addition. The profiles revealed a fast phase, in which about half of the reaction was completed, followed by a slow phase which adds somewhat to the yield, reflecting the incomplete motor assembly and operation identified in the equilibrium experiments. Although further study is needed to fully understand the reasons for the incomplete assembly and operation, this work demonstrates that single-molecule fluorescence, based on its ability to provide detailed in situ structural dynamics information, inaccessible for traditional methods, constitutes an excellent tool for chaperoning the development of DNA-based technology.
- Published
- 2012
10. Disentangling Subpopulations in Single-Molecule FRET and ALEX Experiments with Photon Distribution Analysis
- Author
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Rula Masoud, Eyal Nir, Roman Tsukanov, Miran Liber, Toma E. Tomov, and Noa Plavner
- Subjects
Models, Molecular ,Photons ,Brightness ,Photon ,Chemistry ,Lasers ,Inverted Repeat Sequences ,Statistics as Topic ,Kernel density estimation ,Spectroscopy, Imaging, and Other Techniques ,Biophysics ,Analytical chemistry ,DNA ,Single-molecule FRET ,Förster resonance energy transfer ,Histogram ,Fluorescence Resonance Energy Transfer ,Nucleic Acid Conformation ,DNA origami ,A-DNA ,Biological system - Abstract
Among the advantages of the single-molecule approach when used to study biomolecular structural dynamics and interaction is its ability to distinguish between and independently observe minor subpopulations. In a single-molecule Förster resonance energy transfer (FRET) and alternating laser excitation diffusion experiment, the various populations are apparent in the resultant histograms. However, because histograms are calculated based on the per-burst mean FRET and stoichiometry ratio and not on the internal photon distribution, much of the acquired information is lost, thereby reducing the capabilities of the method. Here we suggest what to our knowledge is a novel statistical analysis tool that significantly enhances these capabilities, and we use it to identify and isolate static and dynamic subpopulations. Based on a kernel density estimator and a proper photon distribution analysis, for each individual burst, we calculate scores that reflect properties of interest. Specifically, we determine the FRET efficiency and brightness ratio distributions and use them to reveal 1), the underlying structure of a two-state DNA-hairpin and a DNA hairpin that is bound to DNA origami; 2), a minor doubly labeled dsDNA subpopulation concealed in a larger singly labeled dsDNA; and 3), functioning DNA origami motors concealed within a larger subpopulation of defective motors. Altogether, these findings demonstrate the usefulness of the proposed approach. The method was developed and tested using simulations, its rationality is described, and a computer algorithm is provided.
- Published
- 2012
11. Aligning the μs-ALEX Setup
- Author
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Eyal Nir, Mike Heilemann, Shimon Weiss, Devdoot Majumdar, and Achillefs N. Kapanidis
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Materials science ,business.industry ,Protein Conformation ,Lasers ,Clinical Sciences ,Proteins ,MOLECULAR BIOLOGY METHODS ,Laser ,General Biochemistry, Genetics and Molecular Biology ,law.invention ,Förster resonance energy transfer ,law ,Proteins metabolism ,Fluorescence Resonance Energy Transfer ,Optoelectronics ,Biochemistry and Cell Biology ,Sensitivity (control systems) ,business ,Molecular Biology ,Excitation ,Physical Chemistry (incl. Structural) ,Fluorescent Dyes - Abstract
To achieve single-molecule sensitivity and thus have the ability to detect single diffusing fluorophores, careful alignment of the microsecond-alternating laser excitation (μs-Alex) setup is crucial. The following protocol describes routine alignment for 2c-ALEX (532 nm/635 nm) with spectral windows G550–620R650–750.
- Published
- 2015
12. Alternating Laser Excitation for Solution-Based Single-Molecule FRET
- Author
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Eyal Nir, Mike Heilemann, Achillefs N. Kapanidis, Devdoot Majumdar, and Shimon Weiss
- Subjects
chemistry.chemical_classification ,Materials science ,Protein Conformation ,Biomolecule ,Lasers ,Clinical Sciences ,Sorting ,Proteins ,Single-molecule FRET ,Laser ,Fluorescence ,General Biochemistry, Genetics and Molecular Biology ,law.invention ,Förster resonance energy transfer ,chemistry ,Chemical physics ,law ,Fluorescence Resonance Energy Transfer ,Molecule ,Biochemistry and Cell Biology ,Molecular Biology ,Excitation ,Fluorescent Dyes ,Physical Chemistry (incl. Structural) - Abstract
Single-molecule fluorescence resonance energy transfer (smFRET) has been widely applied to the study of fluorescently labeled biomolecules on surfaces and in solution. Sorting single molecules based on fluorescent dye stoichiometry provides one with further layers of information and also enables “filtering” of unwanted molecules from the analysis. We accomplish this sorting by using alternating laser excitation (ALEX) in combination with smFRET measurements; here we describe the implementation of these methodologies for the study of biomolecules in solution.
- Published
- 2015
13. Assembling the μs-ALEX Setup
- Author
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Eyal Nir, Achillefs N. Kapanidis, Mike Heilemann, Devdoot Majumdar, and Shimon Weiss
- Subjects
Materials science ,business.industry ,Protein Conformation ,Lasers ,Clinical Sciences ,Proteins ,Laser ,Molecular biology ,General Biochemistry, Genetics and Molecular Biology ,law.invention ,Förster resonance energy transfer ,law ,Fluorescence Resonance Energy Transfer ,Optoelectronics ,Biochemistry and Cell Biology ,business ,Molecular Biology ,Excitation ,Directly modulated laser ,Fluorescent Dyes ,Physical Chemistry (incl. Structural) - Abstract
This protocol describes the construction of a microsecond-alternating laser excitation (μs-ALEX) using two lasers, a green 532-nm acousto-optically modulated laser and a red 635-nm directly modulated laser.
- Published
- 2015
14. Occupation times and Bessel densities
- Author
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Eyal Nir, Yevgeniy Kovchegov, and Nick Meredith
- Subjects
Statistics and Probability ,Laplace transform ,Mathematical analysis ,Markov process ,Reversible process ,Density estimation ,Random walk ,symbols.namesake ,Fourier transform ,Probability theory ,symbols ,Statistics, Probability and Uncertainty ,Bessel function ,Mathematics - Abstract
Consider a Markov process with countably many states. In order to find a one-state occupation time distribution, we use a combination of Fourier and Laplace transforms in the way that allows for the inversion of the Fourier transform. We derive a closed-form expression for the occupation time distribution in the case of a simple continuous-time random walk on Z and represent the one-state occupation density of a reversible process as a mixture of Bessel densities.
- Published
- 2010
15. From No-Dimensions to N-Dimensions with Parametric Point-Clouds
- Author
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Eyal Nir
- Subjects
Engineering ,Engineering drawing ,Theoretical computer science ,business.industry ,Point cloud ,Building and Construction ,Computer Graphics and Computer-Aided Design ,Computer Science Applications ,Parametric design ,Development (topology) ,Generative Design ,business ,Representation (mathematics) ,Parametric statistics - Abstract
This paper presents an innovative approach towards parametric design using point-clouds as design media. Exposing the internal numeric representation of digital models led to the development of parametric point-clouds as design drivers. A parametric point-cloud concept is presented in this paper, exploring its potential application for behavior modeling, generative design and performance-driven design of building envelopes.
- Published
- 2007
16. Nucleosome Core Particle Disassembly and Assembly Kinetics Studied Using Single-Molecule Fluorescence
- Author
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Yaron Berger, Eyal Nir, Toma E. Tomov, Noa Plavner Hazan, Katalin Tóth, Roman Tsukanov, Joerg Langowski, Rula Masoud, and Miran Liber
- Subjects
biology ,Chemistry ,Kinetics ,Biophysics ,Sodium Chloride ,Single-molecule experiment ,Dissociation (chemistry) ,Fluorescence ,Nucleosomes ,Crystallography ,chemistry.chemical_compound ,Xenopus laevis ,Histone ,Förster resonance energy transfer ,biology.protein ,Escherichia coli ,Fluorescence Resonance Energy Transfer ,Nucleosome ,Animals ,Proteins and Nucleic Acids ,Dimerization ,DNA ,Macromolecule - Abstract
The stability of the nucleosome core particle (NCP) is believed to play a major role in regulation of gene expression. To understand the mechanisms that influence NCP stability, we studied stability and dissociation and association kinetics under different histone protein (NCP) and NaCl concentrations using single-pair Förster resonance energy transfer and alternating laser excitation techniques. The method enables distinction between folded, unfolded, and intermediate NCP states and enables measurements at picomolar to nanomolar NCP concentrations where dissociation and association reactions can be directly observed. We reproduced the previously observed nonmonotonic dependence of NCP stability on NaCl concentration, and we suggest that this rather unexpected behavior is a result of interplay between repulsive and attractive forces within positively charged histones and between the histones and the negatively charged DNA. Higher NaCl concentrations decrease the attractive force between the histone proteins and the DNA but also stabilize H2A/H2B histone dimers, and possibly (H3/H4)2 tetramers. An intermediate state in which one DNA arm is unwrapped, previously observed at high NaCl concentrations, is also explained by this salt-induced stabilization. The strong dependence of NCP stability on ion and histone concentrations, and possibly on other charged macromolecules, may play a role in chromosomal morphology.
- Published
- 2015
17. Site-specific labeling of proteins for single-molecule FRET by combining chemical and enzymatic modification
- Author
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Eyal Nir, Shimon Weiss, and Marcus Jäger
- Subjects
Protein Folding ,alternating laser excitation ,single-molecule spectroscopy ,fluorescence-aided molecular sorting ,Glutamine ,fluorescence resonance energy transfer ,Protein Engineering ,Biochemistry ,transglutaminase ,Fluorescence Resonance Energy Transfer ,Cysteine ,Molecular Biology ,Fluorescent Dyes ,Plant Proteins ,chemistry.chemical_classification ,Transglutaminases ,Substrate (chemistry) ,protein labeling ,Single-molecule FRET ,Protein engineering ,N-terminus ,Enzyme ,Förster resonance energy transfer ,chemistry ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,For the Record ,Thermodynamics ,Protein folding ,Peptides - Abstract
An often limiting factor for studying protein folding by single-molecule fluorescence resonance energy transfer (FRET) is the ability to site-specifically introduce a photostable organic FRET donor (D) and a complementary acceptor (A) into a polypeptide chain. Using alternating-laser excitation and chymotrypsin inhibitor 2 as a model, we show that chemical labeling of a unique cysteine, followed by enzymatic modification of a reactive glutamine in an N-terminally appended substrate sequence recognition tag for transglutaminase (TGase) affords stoichiometrically D-/A-labeled protein suitable for single-molecule FRET experiments. Thermodynamic data indicate that neither the presence of the TGase tag nor D/A labeling perturbs protein stability. As the N terminus in proteins is typically solvent accessible, a TGase tag can (in principle) be appended to any protein of interest by genetic engineering. Two-step chemical/enzymatic labeling may thus represent a simple, low-cost, and widely available strategy for D/A labeling of proteins for FRET-based single-molecule protein folding studies, even for non-protein-experts laboratories.
- Published
- 2006
18. Photochemical selectivity in guanine–cytosine base-pair structures
- Author
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Louis Grace, Pavel Hobza, Eyal Nir, Mattanjah S. de Vries, Martin Kabeláč, and Ali Abo-Riziq
- Subjects
chemistry.chemical_classification ,Guanine ,Multidisciplinary ,Spectrophotometry, Infrared ,Photochemistry ,Chemistry ,Base pair ,Hydrogen bond ,Biomolecule ,Hydrogen Bonding ,Cytosine ,chemistry.chemical_compound ,Ab initio quantum chemistry methods ,Nucleic Acids ,Physical Sciences ,Spectrophotometry, Ultraviolet ,Absorption (electromagnetic radiation) ,Spectroscopy ,Base Pairing - Abstract
Prebiotic chemistry presumably took place before formation of an oxygen-rich atmosphere and thus under conditions of intense short wavelength UV irradiation. Therefore, the UV photochemical stability of the molecular building blocks of life may have been an important selective factor in determining the eventual chemical makeup of critical biomolecules. To investigate the role of UV irradiation in base-pairing we have studied guanine (G) and cytosine (C) base pairs in the absence of the RNA backbone. We distinguished base-pair structures by IR–UV hole-burning spectroscopy as well as by high-level correlated ab initio calculations. The Watson–Crick structure exhibits broad UV absorption, in stark contrast to other GC structures and other base-pair structures. This broad absorption may be explained by a rapid internal conversion that makes this specific base pair arrangement uniquely photochemically stable.
- Published
- 2004
19. Properties of isolated DNA bases, base pairs and nucleosides examined by laser spectroscopy
- Author
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Ch. Plützer, Eyal Nir, M.S. de Vries, and Karl Kleinermanns
- Subjects
Crystallography ,chemistry.chemical_compound ,Hydrogen bond ,Chemistry ,Vibronic spectroscopy ,Infrared spectroscopy ,Keto–enol tautomerism ,Resonance (chemistry) ,Tautomer ,Enol ,Atomic and Molecular Physics, and Optics ,Nucleobase - Abstract
The vibronic spectra of laser desorbed and jet cooled guanine (G) adenine (A), and cytosine (C) consist of bands from four, two and two major tautomers respectively, as revealed by UV-UV and IR-UV double resonance spectroscopy. The vibronic spectrum of adenine around 277 nm consists of weak nπ* and strong ππ* transitions, based on IR-UV and deuteration experiments. Precise ionization potentials of G and A were determined with 2-color, 2-photon ionization. We also measured vibronic and IR spectra of several base pairs. GC exhibits a HNH ... OH/NH ... N/C=O ... HNH bonding similar to the Watson-Crick GC base pair but with C as enol tautomer. One GG isomer exhibits non-symmetric hydrogen bonding with HNH ... N/NH ... N/C=O ... HNH interactions. A second observed GG isomer has a symmetrical hydrogen bond arrangement with C=O ... NH/NH ... O=C bonding. Two CC isomers were observed with symmetrical C=O ... NH/NH ... O=C bonding and nonsymmetrical C=O ... HNH/NH ... N interaction, respectively. Guanosine (Gs), 2-DeoxyGs und 3-DeoxyGs each exhibit only one isomer in the investigated wavelength range around 290 nm with a strong intramolecular sugar(5-OH) ... enolguanine(3-N) hydrogen bond.
- Published
- 2002
20. REMPI spectroscopy of cytosine
- Author
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M. Müller, Eyal Nir, M.S. de Vries, and Louis Grace
- Subjects
chemistry.chemical_compound ,Pyrimidine ,chemistry ,Ionization ,General Physics and Astronomy ,Vibronic spectroscopy ,Keto–enol tautomerism ,Photoionization ,Physical and Theoretical Chemistry ,Photochemistry ,Spectroscopy ,Spectral line ,Cytosine - Abstract
We report resonant two-photon ionization spectra of laser desorbed, jet cooled, cytosine, 1-methyl cytosine, 5-methyl cytosine, and dimers of these. Unlike other pyrimidine bases, cytosine exhibits vibronic spectra with sharp features in two spectral regions, separated by about 5000 cm 1 . We interpret these as being due to two tautomeric forms, one keto and one enol. The dimers absorb at wavelengths that are intermediate between those of the two monomer forms. By UV–UV hole burning we determined the numbers of isomers contributing to each spectrum and by delayed two color ionization we determined triplet lifetimes. We observed hydrogen transfer between bases both in collisions between monomers and after photo-excitation in clusters. 2002 Published by Elsevier Science B.V.
- Published
- 2002
21. Pairing of the nucleobases guanine and cytosine in the gas phase studied by IR–UV double-resonance spectroscopy and ab initio calculations
- Author
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Karl Kleinermanns, Petra Imhof, Eyal Nir, M.S. de Vries, and Ch. Janzen
- Subjects
Dimer ,General Physics and Astronomy ,Infrared spectroscopy ,Photochemistry ,Resonance (chemistry) ,Tautomer ,Enol ,Nucleobase ,chemistry.chemical_compound ,Crystallography ,chemistry ,Ab initio quantum chemistry methods ,Physical and Theoretical Chemistry ,Spectroscopy - Abstract
We present R2PI, IR–UV and UV–UV double resonance measurements of the guanine–cytosine (G–C) dimer formed in a supersonic jet. We show that there is only one isomer of G–C in the investigated wavelength range from 33200 to 34100 cm−1. We assigned the observed G–C isomer to a specific structure, based on comparisons of the IR spectra of the G and C monomers with the G–C dimer in the range of the OH and NH stretching vibrations and ab initio-calculated vibrational frequencies and dimer stabilities. The cluster exhibits an HNH⋯O/NH⋯N/CO⋯HNH bonding similar to the Watson–Crick G–C base pair bonding but with C as the enol tautomer. We did not observe any keto–keto or enol–enol G–C dimers in the investigated wavelength region.
- Published
- 2002
22. Pairing of the nucleobase guanine studied by IR–UV double-resonance spectroscopy and ab initio calculations
- Author
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Petra Imhof, Mattanjah S. de Vries, Eyal Nir, Karl Kleinermanns, and Christoph Janzen
- Subjects
Hydrogen bond ,Chemistry ,Dimer ,Ab initio ,General Physics and Astronomy ,Infrared spectroscopy ,Resonance (chemistry) ,Photochemistry ,Tautomer ,Enol ,Crystallography ,chemistry.chemical_compound ,Ab initio quantum chemistry methods ,Physical and Theoretical Chemistry - Abstract
In this paper we present detailed R2PI spectra with IR–UV and UV–UV double resonance measurements of the guanine dimer (GG) and its methylated derivatives. We show that there are two isomers of GG in the investigated wavelength range from 32565 to 33600 cm−1. We were able to assign the two isomers to specific structures based on comparison of the intermolecular vibronic patterns of the dimers with and without methylation, on analysis of the IR spectra in the range of the OH and NH stretching vibrations and on comparison with ab initio calculated dimer stabilities and vibrational frequencies. In both structures both guanine moieties are in the keto tautomeric form, even though the enol tautomers are also present in the beam. One isomer exhibits nonsymmetric hydrogen bonding with HNH⋯N, NH⋯N and CO⋯HNH interactions (K9K7-2). The other isomer has a symmetrical hydrogen bond arrangement with CO⋯NH/NH⋯OC bonding (K9K7-1). The most stable guanine dimer forms CO⋯NH/NH⋯OC hydrogen bonds and has C2h symmetry (K9K9-1). Due to its strong exciton splitting the allowed S0–S2 transition is outside the investigated spectral range. We did not observe any keto–enol or enol–enol dimers in the investigated wavelength region. The calculations predict these dimers to be considerably less stable.
- Published
- 2002
23. A bipedal DNA motor that travels back and forth between two DNA origami tiles
- Author
-
Yaron Berger, Toma E. Tomov, Roman Tsukanov, Miran Liber, and Eyal Nir
- Subjects
Computer science ,Nanotechnology ,General Chemistry ,DNA ,Topology ,Molecular machine ,Biomechanical Phenomena ,Biomaterials ,chemistry.chemical_compound ,Successful operation ,chemistry ,visual_art ,DNA nanotechnology ,Molecular motor ,visual_art.visual_art_medium ,DNA origami ,General Materials Science ,Tile ,Dimerization ,Biotechnology - Abstract
In this work, the successful operation of a dynamic DNA device constructed from two DNA origami building blocks is reported. The device includes a bipedal walker that strides back and forth between the two origami tiles. Two different DNA origami tiles are first prepared separately; they are then joined together in a controlled manner by a set of DNA strands to form a stable track in high yield as confirmed by single-molecule fluorescence (SMF). Second, a bipedal DNA motor, initially attached to one of the two origami units and operated by sequential interaction with "fuel" and "antifuel" DNA strands, moves from one origami tile to another and then back again. The operational yield, measured by SMF, was similar to that of a motor operating on a similar track embedded in a single origami tile, confirming that the transfer across the junction from one tile to the other does not result in dissociation that is any more than that of steps on a single tile. These results demonstrate that moving parts can reliably travel from one origami unit to another, and it demonstrates the feasibility of dynamic DNA molecular machines that are made of more than a single origami building block. This study is a step toward the development of motors that can stride over micrometer distances.
- Published
- 2014
24. IR–UV double-resonance spectroscopy of the nucleobase adenine
- Author
-
M.S. de Vries, Karl Kleinermanns, Eyal Nir, and Chr. Plützer
- Subjects
Chemistry ,Ab initio ,General Physics and Astronomy ,Infrared spectroscopy ,Resonance ,Molecule ,Physical and Theoretical Chemistry ,Photochemistry ,Spectroscopy ,Tautomer ,Spectral line ,Nucleobase - Abstract
We present R2PI and IR–UV double resonance spectra of the nucleobase adenine seeded in a supersonic jet. We show that there is only one tautomer of adenine which absorbs in the wavelength range 36 050 to 36 700 cm−1. The IR spectra, measured in the range 3200 to 3700 cm−1, show bands at 3452, 3508 and 3569 cm−1, which we assign to the symmetric NH2 , N–H and antisymmetric NH2 stretching vibrations of a single tautomer of adenine. We compare the experimental IR–UV double resonance spectra with ab initio based normal mode calculations. The observed tautomer is most probably the 9H amino-form of adenine.
- Published
- 2001
25. Guanine tautomerism revealed by UV–UV and IR–UV hole burning spectroscopy
- Author
-
Eyal Nir, Ch. Janzen, K. Kleinermanns, Petra Imhof, and M.S. de Vries
- Subjects
Guanine ,Ab initio ,General Physics and Astronomy ,Resonance ,Infrared spectroscopy ,Laser ,Photochemistry ,Tautomer ,Spectral line ,law.invention ,chemistry.chemical_compound ,chemistry ,law ,Physical and Theoretical Chemistry ,Spectroscopy - Abstract
The vibronic spectrum of laser desorbed and jet cooled guanine consists of bands from three different tautomers of guanine as revealed by UV–UV and IR–UV double resonance spectroscopy. 1-methylguanine, in which the Keto–Enol tautomerism is blocked, shows hole burning spectra from the 9H-and 7H-Keto form. A comparison of the vibronic pattern of the different tautomers demonstrates that the vibronic spectrum built on the redmost guanine band at 32 870 cm−1 (electronic origin 0) can be traced back to the 9H-Enol tautomer, while the spectra built on the origins at 0+404 cm−1 and 0+1044 cm−1 stem from the two Keto tautomers. The IR–UV double resonance spectra of the OH-and NH-stretch vibrations of the different tautomers support this assignment. The UV and IR spectra can be partly assigned by comparison with ab initio calculated vibrational frequencies and with the help of deuteration experiments.
- Published
- 2001
26. On the Photochemistry of Purine Nucleobases
- Author
-
K. Kleinermanns, Eyal Nir, Mattanjah S. de Vries, Louis Grace, and Elektrochemie I
- Subjects
Purine ,chemistry.chemical_compound ,chemistry ,Computational chemistry ,Guanine ,Guanosine ,Physical and Theoretical Chemistry ,Related derivatives ,Nucleobase - Abstract
We present R2PI spectra of a series of purine nucleobases, nucleosides, and related derivatives, including adenine, guanine, guanosine, 2-aminopurine, 2,6-diaminopurine, 9-ethyladenine, and 9-methy...
- Published
- 2001
27. Conformational dynamics of DNA hairpins at millisecond resolution obtained from analysis of single-molecule FRET histograms
- Author
-
Yaron Berger, Eyal Nir, Roman Tsukanov, Toma E. Tomov, and Miran Liber
- Subjects
Total internal reflection fluorescence microscope ,Base Sequence ,Resolution (electron density) ,Single-molecule FRET ,DNA ,Surfaces, Coatings and Films ,Folding (chemistry) ,Crystallography ,Transition state theory ,chemistry.chemical_compound ,Förster resonance energy transfer ,chemistry ,Materials Chemistry ,Biophysics ,Fluorescence Resonance Energy Transfer ,DNA origami ,Nucleic Acid Conformation ,Physical and Theoretical Chemistry - Abstract
Here we provide high resolution study of DNA hairpin dynamics achieved by probability distribution analysis (PDA) of diffusion-based single-molecule Förster resonance energy transfer (sm-FRET) histograms. The opening and closing rates of three hairpins both free and attached to DNA origami were determined. The agreement with rates previously obtained using the total internal reflection (TIRF) technique and between free hairpins and hairpins attached to origami validated the PDA and demonstrated that the origami had no influence on the hairpin dynamics. From comparison of rates of four DNA hairpins, differing only in stem sequence, and from comparison with rates calculated using nearest-neighbor method and standard transition state theory, we conclude that the unfolding reaction resembles that of melting of DNA duplex with a corresponding sequence and that the folding reaction depends on counterion concentration and not on stem sequence. Our validation and demonstration of the PDA method will encourage its implementation in future high-resolution dynamic studies of freely diffusing biomolecules.
- Published
- 2013
28. Resonance-Enhanced Multiphoton Ionization Spectroscopy of Dipeptides
- Author
-
Rami Cohen, Eyal Nir, Louis Grace, Mattanjah S. de Vries, and Beth Brauer
- Subjects
chemistry.chemical_classification ,Resonance-enhanced multiphoton ionization ,chemistry ,Stereochemistry ,Ionization ,Molecule ,Phenylalanine ,Physical and Theoretical Chemistry ,Chromophore ,Ring (chemistry) ,Spectroscopy ,Amino acid - Abstract
We report resonance-enhanced multiphoton ionization (REMPI) spectroscopy of laser-desorbed, jet-cooled dipeptides, containing either tyrosine or phenylalanine as an aromatic chromophore (C). The single amino acids have multiple origins that we interpret as arising from two types of conformations, with the carboxyl group either anti or gauche with respect to the ring. Spectra for dipeptides of the form X−C, for example, Ala-Tyr, are similar to those of the corresponding single amino acid. On the other hand, spectra for dipeptides of the form C−X, for example, Tyr-Ala, show significant changes in the peaks, which we associate with gauche conformations. This observation can be understood in terms of an interaction between the carboxyl terminus and the ring, associated with the molecule assuming a folded conformation.
- Published
- 2000
29. Rational design of DNA motors: fuel optimization through single-molecule fluorescence
- Author
-
Toma E. Tomov, Rula Masoud, Miran Liber, Roman Tsukanov, Noa Plavner, and Eyal Nir
- Subjects
Models, Molecular ,Chemistry ,Rational design ,DNA walker ,Nanotechnology ,General Chemistry ,DNA ,Single-molecule experiment ,Biochemistry ,Catalysis ,Molecular machine ,Fluorescence ,Biomechanical Phenomena ,Reaction rate ,chemistry.chemical_compound ,Kinetics ,Colloid and Surface Chemistry ,Yield (chemistry) ,A-DNA ,Biological system - Abstract
While numerous DNA-based molecular machines have been developed in recent years, high operational yield and speed remain a major challenge. To understand the reasons for the limited performance, and to find rational solutions, we applied single-molecule fluorescence techniques and conducted a detailed study of the reactions involved in the operation of a model system comprised of a bipedal DNA walker that strides on a DNA origami track powered by interactions with fuel and antifuel strands. Analysis of the kinetic profiles of the leg-lifting reactions indicates a pseudo-first-order antifuel binding mechanism leading to a rapid and complete leg-lifting, indicating that the fuel-removal reaction is not responsible for the 1% operational yield observed after six steps. Analysis of the leg-placing reactions showed that although increased concentrations of fuel increase the reaction rate, they decrease the yield by consecutively binding the motor and leading to an undesirable trapped state. Recognizing this, we designed asymmetrical hairpin-fuels that by regulating the reaction hierarchy avoid consecutive binding. Motors operating with the improved fuels show 74% yield after 12 consecutive reactions, a dramatic increase over the 1% observed for motors operating with nonhairpin fuels. This work demonstrates that studying the mechanisms of the reactions involved in the operation of DNA-based molecular machines using single-molecule fluorescence can facilitate rationally designed improvements that increase yield and speed and promote the applicability of DNA-based machines.
- Published
- 2013
30. A Meta-Analysis of the Effectiveness of the 'But You Are Free' Compliance-Gaining Technique
- Author
-
Eyal, Nir
- Subjects
Research - Published
- 2013
- Full Text
- View/download PDF
31. Sample Preparation and Data Acquisition for μs-ALEX
- Author
-
Shimon Weiss, Devdoot Majumdar, Mike Heilemann, Achillefs N. Kapanidis, and Eyal Nir
- Subjects
Materials science ,Protein Conformation ,business.industry ,Lasers ,Clinical Sciences ,Proteins ,Laser ,Molecular biology ,General Biochemistry, Genetics and Molecular Biology ,Dilution ,law.invention ,Förster resonance energy transfer ,Data acquisition ,law ,Fluorescence Resonance Energy Transfer ,Optoelectronics ,Sample preparation ,Biochemistry and Cell Biology ,business ,Molecular Biology ,Excitation ,Physical Chemistry (incl. Structural) ,Fluorescent Dyes - Abstract
This protocol describes the preparation of samples and data acquisition for microsecond-alternating laser excitation. Sample preparation requires a dilution that ensures the detection of single events.
- Published
- 2015
32. Fragment-Free Mass Spectrometric Analysis with Jet Cooling/VUV Photoionization
- Author
-
M.S. de Vries, H. E. Hunziker, and Eyal Nir
- Subjects
Chemistry ,Analytical chemistry ,Photoionization ,Laser ,Mass spectrometry ,Analytical Chemistry ,law.invention ,Wavelength ,Fragmentation (mass spectrometry) ,law ,Desorption ,Physics::Atomic and Molecular Clusters ,Mass spectrum ,Molecule ,Physics::Chemical Physics - Abstract
We show that it is possible to obtain fragment-free mass spectra of large molecules by a combination of laser desorption, jet cooling, and VUV single-photon photoionization. The ability to obtain parent molecular masses is particularly important for the analysis of mixtures, such as combinations of fully saturated hydrocarbons. By varying the cooling conditions, we can also achieve partial fragmentation in order to obtain further structural information. The use of different wavelengths provides additional selectivity between aromatic and aliphatic compounds.
- Published
- 2011
33. Single-molecule FRET reveals sugar-induced conformational dynamics in LacY
- Author
-
Eyal Nir, Vladimir N. Kasho, Devdoot Majumdar, Irina Smirnova, H. Ronald Kaback, Xiangxu Kong, and Shimon Weiss
- Subjects
Lactose permease ,Models, Molecular ,spectroscopy ,Cytoplasm ,major facilitator superfamily ,Mutant ,Carbohydrates ,transporters ,Ligands ,lactose permease ,Fluorescence Resonance Energy Transfer ,Single-molecule Chemistry and Biology Special Feature ,Multidisciplinary ,Chemistry ,Membrane Transport Proteins ,Single-molecule FRET ,Periplasmic space ,Major facilitator superfamily ,Protein Structure, Tertiary ,Crystallography ,Förster resonance energy transfer ,Symporter ,Mutation ,bacteria ,Carbohydrate Metabolism ,single-molecule ,Protein Binding - Abstract
The N- and C-terminal six-helix bundles of lactose permease (LacY) form a large internal cavity open on the cytoplasmic side and closed on the periplasmic side with a single sugar-binding site at the apex of the cavity near the middle of the molecule. During sugar/H + symport, an outward-facing cavity is thought to open with closing of the inward-facing cavity so that the sugar-binding site is alternately accessible to either face of the membrane. In this communication, single-molecule fluorescence (Förster) resonance energy transfer is used to test this model with wild-type LacY and a conformationally restricted mutant. Pairs of Cys residues at the ends of two helices on the cytoplasmic or periplasmic sides of wild-type LacY and the mutant were labeled with appropriate donor and acceptor fluorophores, single-molecule fluorescence resonance energy transfer was determined in the absence and presence of sugar, and distance changes were calculated. With wild-type LacY, binding of a galactopyranoside, but not a glucopyranoside, results in a decrease in distance on the cytoplasmic side and an increase in distance on the periplasmic side. In contrast, with the mutant, a more pronounced decrease in distance and in distance distribution is observed on the cytoplasmic side, but there is no change on the periplasmic side. The results are consistent with the alternating access model and indicate that the defect in the mutant is due to impaired ligand-induced flexibility on the periplasmic side.
- Published
- 2007
34. Alternating laser excitation spectroscopy of freely diffusing single molecule: Applications to bimolecular structure, dynamics and interactions
- Author
-
Eyal Nir, Marcus Jäger, and Shimon Weiss
- Subjects
Excitation spectroscopy ,Chemistry ,Chemical physics ,law ,Dynamics (mechanics) ,Structure (category theory) ,Molecule ,Physical chemistry ,Laser ,law.invention - Published
- 2007
35. Introducing improved structural properties and salt dependence into a coarse-grained model of DNA
- Author
-
Ferdinando Randisi, Thomas E. Ouldridge, Petr Šulc, Majid Mosayebi, Eyal Nir, Roman Tsukanov, Flavio Romano, John S. Schreck, Jonathan P. K. Doye, Ard A. Louis, and Benedict E. K. Snodin
- Subjects
Materials science ,Static Electricity ,Stacking ,DNA ,Elasticity ,Fluorescence Resonance Energy Transfer ,Molecular Dynamics Simulation ,Nucleic Acid Conformation ,Salts ,Thermodynamics ,Transition Temperature ,Models, Genetic ,Physics and Astronomy (all) ,Physical and Theoretical Chemistry ,FOS: Physical sciences ,General Physics and Astronomy ,02 engineering and technology ,Condensed Matter - Soft Condensed Matter ,010402 general chemistry ,01 natural sciences ,Settore FIS/03 - Fisica della Materia ,chemistry.chemical_compound ,Genetic ,Stack (abstract data type) ,Models ,DNA origami ,Physics - Biological Physics ,Molecular biophysics ,Biomolecules (q-bio.BM) ,021001 nanoscience & nanotechnology ,Electrostatics ,Settore FIS/02 - Fisica Teorica, Modelli e Metodi Matematici ,0104 chemical sciences ,Thymine ,Range (mathematics) ,Quantitative Biology - Biomolecules ,chemistry ,Biological Physics (physics.bio-ph) ,FOS: Biological sciences ,Soft Condensed Matter (cond-mat.soft) ,0210 nano-technology ,Biological system - Abstract
We introduce an extended version of oxDNA, a coarse-grained model of deoxyribonucleic acid (DNA) designed to capture the thermodynamic, structural, and mechanical properties of single- and double-stranded DNA. By including explicit major and minor grooves and by slightly modifying the coaxial stacking and backbone-backbone interactions, we improve the ability of the model to treat large (kilobase-pair) structures, such as DNA origami, which are sensitive to these geometric features. Further, we extend the model, which was previously parameterised to just one salt concentration ([Na(+)] = 0.5M), so that it can be used for a range of salt concentrations including those corresponding to physiological conditions. Finally, we use new experimental data to parameterise the oxDNA potential so that consecutive adenine bases stack with a different strength to consecutive thymine bases, a feature which allows a more accurate treatment of systems where the flexibility of single-stranded regions is important. We illustrate the new possibilities opened up by the updated model, oxDNA2, by presenting results from simulations of the structure of large DNA objects and by using the model to investigate some salt-dependent properties of DNA.
- Published
- 2015
36. DNA Motors: A Bipedal DNA Motor that Travels Back and Forth between Two DNA Origami Tiles (Small 5/2015)
- Author
-
Yaron Berger, Roman Tsukanov, Eyal Nir, Toma E. Tomov, and Miran Liber
- Subjects
Nanotechnology ,General Chemistry ,Biology ,Single-molecule experiment ,Molecular machine ,Biomaterials ,chemistry.chemical_compound ,chemistry ,DNA nanotechnology ,Molecular motor ,Biophysics ,DNA origami ,General Materials Science ,DNA ,Biotechnology - Published
- 2015
37. On Design Continuity with Smart Cloud of Points
- Author
-
Eyal Nir and Guedi Capeluto
- Published
- 2006
38. Using Histogram Matrices as an Interface for Designing with Parametric Point-Clouds
- Author
-
Eyal Nir
- Published
- 2006
39. Smart Cloud-of-Points Model: Point-based Digital Media and Tools for Architectural Design
- Author
-
Eyal Nir and Guedi Capeluto
- Published
- 2005
40. Conformers of guanosines and their vibrations in the electronic ground and excited states, as revealed by double-resonance spectroscopy and ab initio calculations
- Author
-
Eyal Nir, Mattanjah S. de Vries, Karl Kleinermanns, and Isabel Hünig
- Subjects
Models, Molecular ,Guanosine ,Chemistry ,Electron Spin Resonance Spectroscopy ,Infrared spectroscopy ,Tautomer ,Vibration ,Atomic and Molecular Physics, and Optics ,Crystallography ,Computational chemistry ,Ab initio quantum chemistry methods ,Spectrophotometry ,Excited state ,Spectral hole burning ,Vibronic spectroscopy ,Nucleic Acid Conformation ,Quantum Theory ,Physical and Theoretical Chemistry ,Spectroscopy ,Conformational isomerism ,Mathematics - Abstract
two-photon ionization. The results showed that nucleobases and complexes composed of paired bases can be studied in great detail in the absence of the external effects of solvent interactions and the collective modes of DNA. Here we show by means of double-resonance laser spectroscopy and ab initio calculations that, in the investigated wavelength range around 290 nm, guanosine (Gs), 2-deoxyguanosine (2-deoxyGs), and 3-deoxyguanosine (3-deoxyGs) each exhibit only the spectrum of one stable isomer, which is stabilized by a strong intramolecular sugar(5-OH) ¥¥¥ enolguanine(3-N) hydrogen bond. Figure 1 shows a possible structure of guanosine (Gs) with guanine in its enol form; atoms which are important for the following discussion are numbered. The R2PI spectra of laser-desorbed Gs, 2-deoxyGs, 3 ± deoxyGs have already been published. [5, 21] In the earlier paper we speculated that two conformers might contribute to the vibronic spectra of Gs and 3deoxyGs. To test this hypothesis we performed UV ± UV spectral hole burning (SHB) measurements. Figure 2 shows that each of the three spectra originates from only one conformer, because every single peak in the R2PI spectrum is reflected in the corresponding SHB spectrum. For comparison the R2PI spectrum of 2,3-diisopropylguanosine is shown as well. Figure 3 shows the IR spectra obtained from IR ± UV measurements with the UV laser fixed to the most intense vibronic transition of the respective guanosine, indicated by asterisks in Figure 2. For comparison the IR spectra of 2,3-diisopropylGs and one enol and one keto guanine (G) tautomer are also shown. In 2,3-diisopropylGs, the OH groups of the sugar moiety in the 2and 3-positions are blocked. Therefore, its IR spectrum does not exhibit the corresponding OH vibrations but agrees well with
- Published
- 2004
41. Pairing of isolated nucleobases: double resonance laser spectroscopy of adenine-thymine
- Author
-
Karl Kleinermanns, Eyal Nir, Isabel Hünig, Mattanjah S. de Vries, and Christian Plützer
- Subjects
Photons ,Spectrophotometry, Infrared ,Hydrogen bond ,Chemistry ,Ultraviolet Rays ,Adenine ,Analytical chemistry ,Ab initio ,Infrared spectroscopy ,Hydrogen Bonding ,Resonance (chemistry) ,Methylation ,Atomic and Molecular Physics, and Optics ,Nucleobase ,Thymine ,Crystallography ,chemistry.chemical_compound ,Models, Chemical ,Ab initio quantum chemistry methods ,Spectrophotometry ,Physics::Atomic and Molecular Clusters ,Physics::Chemical Physics ,Physical and Theoretical Chemistry ,Spectroscopy - Abstract
The vibronic spectrum of the adenine ± thymine (A ± T) base pair was obtained byone-color resonant two-photon ionization (R2PI) spectroscopyin a free jet of thermallyevaporated A and T under conditions favorable for formation of small clusters. The onset of the spectrum at 35 064 cm 1 exhibits a large red shift relative to the ±* origin of 9H-adenine at 36 105 cm 1 . The IR ± UV spectrum was assigned to cluster structures with HNH ¥¥¥ OC/N ¥¥¥ HN hydrogen bonding bycomparison with the IR spectra of A and T monomers and with ab initio calculated vibrational spectra of the most stable A ± T isomers. The Watson ± Crick A ± T base pair is not the most stable base-pair structure at different levels of ab initio theory, and its vibrational spectrum is not in agreement with the observed experimental spectrum. Experiments with methylated A and T were performed to further support the structural assignment.
- Published
- 2003
42. Pairing of isolated nucleic-acid bases in the absence of the DNA backbone
- Author
-
Eyal Nir, Karl Kleinermanns, and Mattanjah S. de Vries
- Subjects
chemistry.chemical_classification ,Multidisciplinary ,Guanine ,Hydrogen bond ,Base pair ,DNA damage ,DNA replication ,DNA ,Crystallography ,chemistry.chemical_compound ,Cytosine ,chemistry ,Biochemistry ,Nucleic acid ,Electrochemistry ,Molecule ,Nucleotide ,Gases ,Base Pairing - Abstract
The two intertwined strands of DNA are held together through base pairing—the formation of hydrogen bonds between bases located opposite each other on the two strands. DNA replication and transcription involve the breaking and re-forming of these hydrogen bonds, but it is difficult to probe these processes directly. For example, conventional DNA spectroscopy1,2,3 is dominated by solvent interactions, crystal modes and collective modes of the DNA backbone; gas-phase studies, in contrast, can in principle measure interactions between individual molecules in the absence of external effects, but require the vaporization of the interacting species without thermal degradation4,5,6,7,8,9. Here we report the generation of gas-phase complexes comprising paired bases, and the spectroscopic characterization of the hydrogen bonding in isolated guanine–cytosine (G–C) and guanine–guanine (G–G) base pairs. We find that the gas-phase G–C base pair adopts a single configuration, which may be Watson–Crick, whereas G–G exists in two different configurations, and we see evidence for proton transfer in the G–C pair, an important step in radiation-induced DNA damage pathways10. Interactions between different bases and between bases and water molecules can also be characterized by our approach, providing stringent tests for high-level ab initio computations that aim to elucidate the fundamental aspects of nucleotide interactions11,12,13.
- Published
- 2001
43. REMPI Spectroscopy of Laser Desorbed Guanosines
- Author
-
Eyal Nir, Petra Imhof, Mattanjah S. de Vries, and K. Kleinermanns
- Subjects
Colloid and Surface Chemistry ,law ,Chemistry ,Analytical chemistry ,General Chemistry ,Spectroscopy ,Laser ,Biochemistry ,Catalysis ,law.invention - Published
- 2000
44. REMPI Spectroscopy of Jet-Cooled Guanine
- Author
-
Mattanjah S. de Vries, and Beth Brauer, Eyal Nir, and Louis Grace
- Subjects
Jet (fluid) ,Range (particle radiation) ,Guanine ,Analytical chemistry ,General Chemistry ,Biochemistry ,Fluence ,Catalysis ,chemistry.chemical_compound ,Colloid and Surface Chemistry ,chemistry ,Ionization ,Atomic physics ,Spectroscopy - Abstract
Figure 1. REMPI spectrum of jet-cooled guanine. The spectrum consists of four parts, as indicated by the horizontal arrows, that were recorded independently. In each part the ionization laser fluence varies such that it is highest in the center of the respective range. The peaks are not normalized for laser fluence. The top panel shows IR data from ref 8, plotted relative to the lowest energy peak in the REMPI spectrum for comparison.
- Published
- 1999
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