Search

The ataxia telangiectasia–mutated and Rad3-related (ATR) kinase is a master regulator of DNA damage response and replication stress in humans, but the mechanism of its activation remains unclear. ATR acts together with its partner ATRIP. Using cryo–electron microscopy, we determined the structure of intact Mec1-Ddc2 (the yeast homolog of ATR-ATRIP),which is poised for catalysis, at a resolution of 3.9 angstroms.Mec1-Ddc2 forms a dimer of heterodimers through the PRD and FATdomains of Mec1 and the coiled-coil domain of Ddc2. The PRD and Bridge domains inMec1 constitute critical regulatory sites. The activation loop ofMec1 is inhibited by the PRD, revealing an allostericmechanism of kinase activation. Our study clarifies the architecture of ATR-ATRIP and provides a structural framework for the understanding of ATR regulation. [ABSTRACT FROM AUTHOR]

A cloud-to-ground lightning flash with nine return strokes has been recorded using a high speed slitless spectrograph and a system composed of a fast antenna and a slow antenna. Based on the spectral data and the synchronous electric field changes that were caused by the lightning, the electrical conductivity, the channel radii, the resistance per unit length, the peak current, the thermal power at the instant of peak current, and the heat energy per unit length during the first 5 is in the discharge channel have all been calculated. The results indicate that the channel radii have linear relationships with the peak current. The thermal power at the peak current time increases with increasing resistance, but exponential decays with the square of the peak current. [ABSTRACT FROM AUTHOR]

c-Cbl-associated protein (CAP) is an important cytoskeletal adaptor protein involved in the regulation of adhesion turnover. The interaction between CAP and vinculin is critical for the recruitment of CAP to focal adhesions. The tandem SH3 domains (herein termed SH3a and SH3b) of CAP are responsible for its interaction with vinculin. However, the structural mechanism underlying the interaction between CAP and vinculin is poorly understood. In this manuscript, we report the solution structure of the tandem SH3 domains of CAP. Our NMR and ITC data indicate that the SH3a and SH3b domains of CAP simultaneously bind to a long proline-rich region of vinculin with different binding specificities. Furthermore, the crystal structures of the individual SH3a and SH3b domains complexed with their substrate peptides indicate that Q807SH3a and D881SH3b are the critical residues determining the different binding specificities of the SH3 domains. Based on the obtained structural information, a model of the SH3ab-vinculin complex was generated using MD simulation and SAXS data. [ABSTRACT FROM AUTHOR]

Using the spectra of a cloud-to-ground (CG) lightning flash with multiple return strokes and combining with the synchronous radiated electrical field information, the linear charge density, the channel radius, the energy per unit length, the thermal energy, and the energy of dissociation and ionization in discharge channel are calculated with the aid of an electrodynamic model of lightning. The conclusion that the initial radius of discharge channel is determined by the duration of the discharge current is confirmed. Moreover, the correlativity of several parameters has been analyzed first. The results indicate that the total intensity of spectra is positive correlated to the channel initial radius. The ionization and thermal energies have a linear relationship, and the dissociation energy is correlated positively to the ionization and thermal energies, the energy per unit length is in direct proportion to the square of initial radius in different strokes of one CG lightning. [ABSTRACT FROM AUTHOR]

Purpose: Recent preclinical and clinical studies show that dyes that excite and fluoresce in the near-infrared range may be used for tracking and detecting disease targets in vivo. A method for quantifying free dye molecules in antibody conjugate preparations is required for agent batch release and for translation into the clinic.Procedures: Herein, we developed and validated a SDS-PAGE method to determine the percentage of free IRDye 800 CW in (DTPA)(n)-trastuzumab-(IRDye 800)(m) conjugate sample preparations in which high-performance liquid chromatography (HPLC) assessment of free dye was not possible.Results: The SDS-PAGE assay was accurate and valid for free IRDye 800 CW amounts between 38 and 4 mol% of total dye. Gel sample preparation reagent affected the specificity of the assay, and lower and upper limits of quantitation and detection were determined.Conclusion: This method may be applicable to other near-infrared dye-conjugated antibody-based imaging agents in which HPLC assessment of purity is not feasible. This validated method for quality assurance will facilitate the translation of dual-labeled antibody conjugates for nuclear and optical imaging. [ABSTRACT FROM AUTHOR]

Cloud-to-ground lightning with six return strokes has been recorded by slitless spectrograph and the system of fast antenna and slow antenna. The physical parameters of the discharge channel have been obtained based on the combination of spectra and synchronous radiated electric field. The resistance and internal electric field of the channel are studied as the focus in this paper. The results show that the resistances per unit length of the lightning channel are in the order of 10-2-10-1Ω/m and the internal electric field strengths are in the order of 10³V/m. [ABSTRACT FROM AUTHOR]

Ni-W-Cr-P alloy coatings were prepared by electroless deposition on stainless steel. The effects of heat-treatment on the structure and phase transformation behavior, microhardness of the Ni-W-Cr-P alloy coatings were mainly investigated. XRD analysis shows that as-deposited Ni-W-Cr-P coatings were microcrystalline; the precipitation of Ni3P was observed at the annealing temperature of 400°C; after heat-treatment at 500°C the crystallization of Ni3P and Ni was near completion; at 600°C new phase Cr1.12Ni2.88 was observed and Ni3P began to decompose. Cr1.07Fe18.93 and Ni17W3 were formed when heated at 700°C, and Ni3P was not found. With increasing temperature to 800°C, FeNi2P and Cr4Ni15W were the only two dominant phases. The experimental results reveal that with an increase in the annealing temperature, micro-hardness of the Ni-W-Cr-P alloy coatings increased, reached the maximum value at 700°C, and then decreased slightly. Annealing temperature dependence of the microhardness and corrosion resistance of the coatings was also observed. The related strengthening mechanism in the electroless deposited Ni-W-Cr-P alloy coatings is also discussed. [ABSTRACT FROM AUTHOR]

Diethylenetriamine-pentaacetic acid (DTPA)-coupled minigastrins are unsuitable for therapeutic application with the available β-emitting radiometals due to low complex stability. Low tumour-to-kidney ratio of the known radiopharmaceuticals is further limiting their potency. We used macrocyclic chelators for coupling to increase complex stability, modified the peptide sequence to enhance radiolytic stability and studied tumour-to-kidney ratio and metabolic stability using 111In-labelled derivatives. Gastrin derivatives with decreasing numbers of glutamic acids were synthesised using 111In as surrogate for therapeutic radiometals for in vitro and in vivo studies. Gastrin receptor affinities of the natIn-metallated compounds were determined by receptor autoradiography using 125I-CCK as radioligand. Internalisation was evaluated in AR4-2J cells. Enzymatic stability was determined by incubating the 111In-labelled peptides in human serum. Biodistribution was performed in AR4-2J-bearing Lewis rats. IC50 values of the natIn-metallated gastrin derivatives vary between 1.2 and 4.8 nmol/L for all methionine-containing derivatives. Replacement of methionine by norleucine, isoleucine, methionine-sulfoxide and methionine-sulfone resulted in significant decrease of receptor affinity (IC50 between 9.9 and 1,195 nmol/L). All cholecystokinin receptor affinities were >100 nmol/L. All 111In-labelled radiopeptides showed receptor-specific internalisation. Serum mean-life times varied between 2.0 and 72.6 h, positively correlating with the number of Glu residues. All 111In-labelled macrocyclic chelator conjugates showed higher tumour-to-kidney ratios after 24 h (0.37–0.99) compared to 111In-DTPA-minigastrin 0 (0.05). Tumour wash out between 4 and 24 h was low. Imaging studies confirmed receptor-specific blocking of the tumour uptake. Reducing the number of glutamates increased tumour-to-kidney ratio but resulted in lower metabolic stability. The properties of the macrocyclic chelator-bearing derivatives make them potentially suitable for clinical purposes. [ABSTRACT FROM AUTHOR]

The article focuses on a study according to which all Ribonuclease P (RNase P) ribozymes share a substrateinduced catalytic mechanism of pre-RNA processing. It mentions that RNase P is a ribonucleoprotein complex, composed of a single catalytic RNA components. It states that the Rpr1 RNA adopts an extended single-layered conformation that maintains a central helical core but lacks longrange RNA-RNA interactions that are essential for structural stability in bacterial RNase P.