775 results on '"Hu, Yilin"'
Search Results
2. Heterologous expression of a fully active Azotobacter vinelandii nitrogenase Fe protein in Escherichia coli.
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Solomon, Joseph, Liu, Yiling, Górecki, Kamil, Quechol, Robert, Lee, Chi, Jasniewski, Andrew, Hu, Yilin, and Ribbe, Markus
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Fe protein ,NifH ,assembly ,heterologous expression ,nitrogenase - Abstract
The functional versatility of the Fe protein, the reductase component of nitrogenase, makes it an appealing target for heterologous expression, which could facilitate future biotechnological adaptations of nitrogenase-based production of valuable chemical commodities. Yet, the heterologous synthesis of a fully active Fe protein of Azotobacter vinelandii (AvNifH) in Escherichia coli has proven to be a challenging task. Here, we report the successful synthesis of a fully active AvNifH protein upon co-expression of this protein with AvIscS/U and AvNifM in E. coli. Our metal, activity, electron paramagnetic resonance, and X-ray absorption spectroscopy/extended X-ray absorption fine structure (EXAFS) data demonstrate that the heterologously expressed AvNifH protein has a high [Fe4S4] cluster content and is fully functional in nitrogenase catalysis and assembly. Moreover, our phylogenetic analyses and structural predictions suggest that AvNifM could serve as a chaperone and assist the maturation of a cluster-replete AvNifH protein. Given the crucial importance of the Fe protein for the functionality of nitrogenase, this work establishes an effective framework for developing a heterologous expression system of the complete, two-component nitrogenase system; additionally, it provides a useful tool for further exploring the intricate biosynthetic mechanism of this structurally unique and functionally important metalloenzyme. IMPORTANCE The heterologous expression of a fully active Azotobacter vinelandii Fe protein (AvNifH) has never been accomplished. Given the functional importance of this protein in nitrogenase catalysis and assembly, the successful expression of AvNifH in Escherichia coli as reported herein supplies a key element for the further development of heterologous expression systems that explore the catalytic versatility of the Fe protein, either on its own or as a key component of nitrogenase, for nitrogenase-based biotechnological applications in the future. Moreover, the clean genetic background of the heterologous expression host allows for an unambiguous assessment of the effect of certain nif-encoded protein factors, such as AvNifM described in this work, in the maturation of AvNifH, highlighting the utility of this heterologous expression system in further advancing our understanding of the complex biosynthetic mechanism of nitrogenase.
- Published
- 2023
3. Heterologous synthesis of the complex homometallic cores of nitrogenase P- and M-clusters in Escherichia coli.
- Author
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Quechol, Robert, Solomon, Joseph, Liu, Yiling, Lee, Chi, Jasniewski, Andrew, Górecki, Kamil, Oyala, Paul, Hedman, Britt, Hodgson, Keith, Hu, Yilin, and Ribbe, Markus
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EPR ,NifB ,NifDK ,XAS ,nitrogenase ,Nitrogenase ,Escherichia coli ,Nitrogen Fixation ,Oxidoreductases ,Azotobacter vinelandii ,Metalloproteins ,Bacterial Proteins - Abstract
Nitrogenase is an active target of heterologous expression because of its importance for areas related to agronomy, energy, and environment. One major hurdle for expressing an active Mo-nitrogenase in Escherichia coli is to generate the complex metalloclusters (P- and M-clusters) within this enzyme, which involves some highly unique bioinorganic chemistry/metalloenzyme biochemistry that is not generally dealt with in the heterologous expression of proteins via synthetic biology; in particular, the heterologous synthesis of the homometallic P-cluster ([Fe8S7]) and M-cluster core (or L-cluster; [Fe8S9C]) on their respective protein scaffolds, which represents two crucial checkpoints along the biosynthetic pathway of a complete nitrogenase, has yet to be demonstrated by biochemical and spectroscopic analyses of purified metalloproteins. Here, we report the heterologous formation of a P-cluster-containing NifDK protein upon coexpression of Azotobacter vinelandii nifD, nifK, nifH, nifM, and nifZ genes, and that of an L-cluster-containing NifB protein upon coexpression of Methanosarcina acetivorans nifB, nifS, and nifU genes alongside the A. vinelandii fdxN gene, in E. coli. Our metal content, activity, EPR, and XAS/EXAFS data provide conclusive evidence for the successful synthesis of P- and L-clusters in a nondiazotrophic host, thereby highlighting the effectiveness of our metallocentric, divide-and-conquer approach that individually tackles the key events of nitrogenase biosynthesis prior to piecing them together into a complete pathway for the heterologous expression of nitrogenase. As such, this work paves the way for the transgenic expression of an active nitrogenase while providing an effective tool for further tackling the biosynthetic mechanism of this important metalloenzyme.
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- 2023
4. mir-605-3p prevents liver premetastatic niche formation by inhibiting angiogenesis via decreasing exosomal nos3 release in gastric cancer
- Author
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Hu, Yilin, Zang, Weijie, Feng, Ying, Mao, Qinsheng, Chen, Junjie, Zhu, Yizhun, and Xue, Wanjiang
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- 2024
- Full Text
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5. Enzymatic Fischer-Tropsch-Type Reactions.
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Lee, Chi, Grosch, Mario, Solomon, Joseph, Weigand, Wolfgang, Hu, Yilin, and Ribbe, Markus
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Nitrogenase ,Hydrocarbons ,Biotechnology - Abstract
The Fischer-Tropsch (FT) process converts a mixture of CO and H2 into liquid hydrocarbons as a major component of the gas-to-liquid technology for the production of synthetic fuels. Contrary to the energy-demanding chemical FT process, the enzymatic FT-type reactions catalyzed by nitrogenase enzymes, their metalloclusters, and synthetic mimics utilize H+ and e- as the reducing equivalents to reduce CO, CO2, and CN- into hydrocarbons under ambient conditions. The C1 chemistry exemplified by these FT-type reactions is underscored by the structural and electronic properties of the nitrogenase-associated metallocenters, and recent studies have pointed to the potential relevance of this reactivity to nitrogenase mechanism, prebiotic chemistry, and biotechnological applications. This review will provide an overview of the features of nitrogenase enzymes and associated metalloclusters, followed by a detailed discussion of the activities of various nitrogenase-derived FT systems and plausible mechanisms of the enzymatic FT reactions, highlighting the versatility of this unique reactivity while providing perspectives onto its mechanistic, evolutionary, and biotechnological implications.
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- 2023
6. Incorporation of an Asymmetric Mo-Fe-S Cluster as an Artificial Cofactor into Nitrogenase.
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Tanifuji, Kazuki, Jasniewski, Andrew, Lee, Chi, Solomon, Joseph, Nagasawa, Takayuki, Ohki, Yasuhiro, Tatsumi, Kazuyuki, Hedman, Britt, Hodgson, Keith, Hu, Yilin, and Ribbe, Markus
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C1 substrate reduction ,artificial enzymes ,hydrocarbons ,nitrogenases ,synthetic Mo−Fe−S clusters ,Hydrocarbons ,Nitrogenase ,Oxidation-Reduction - Abstract
Nitrogenase employs a sophisticated electron transfer system and a Mo-Fe-S-C cofactor, designated the M-cluster [(cit)MoFe7 S9 C]), to reduce atmospheric N2 to bioaccessible NH3 . Previously, we have shown that the cofactor-free form of nitrogenase can be repurposed as a protein scaffold for the incorporation of a synthetic Fe-S cluster [Fe6 S9 (SEt)2 ]4- . Here, we demonstrate the utility of an asymmetric Mo-Fe-S cluster [Cp*MoFe5 S9 (SH)]3- as an alternative artificial cofactor upon incorporation into the cofactor-free nitrogenase scaffold. The resultant semi-artificial enzyme catalytically reduces C2 H2 to C2 H4 , and CN- into short-chain hydrocarbons, yet it is clearly distinct in activity from its [Fe6 S9 (SEt)2 ]4- -reconstituted counterpart, pointing to the possibility to employ molecular design and cluster synthesis strategies to further develop semi-artificial or artificial systems with desired catalytic activities.
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- 2022
7. Radical SAM-dependent formation of a nitrogenase cofactor core on NifB
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Liu, Yiling A, Quechol, Robert, Solomon, Joseph B, Lee, Chi Chung, Ribbe, Markus W, Hu, Yilin, Hedman, Britt, and Hodgson, Keith O
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Chemical Sciences ,Physical Chemistry ,Metalloproteins ,Methyltransferases ,Molybdoferredoxin ,Nitrogenase ,S-Adenosylmethionine ,Biosynthesis ,FeS cluster ,M-cluster ,NifB ,Radical SAM enzyme ,Inorganic Chemistry ,Theoretical and Computational Chemistry ,Other Chemical Sciences ,Inorganic & Nuclear Chemistry ,Inorganic chemistry - Abstract
Nitrogenase is a versatile metalloenzyme that reduces N2, CO and CO2 at its cofactor site. Designated the M-cluster, this complex cofactor has a composition of [(R-homocitrate)MoFe7S9C], and it is assembled through the generation of a unique [Fe8S9C] core prior to the insertion of Mo and homocitrate. NifB is a radical S-adenosyl-L-methionine (SAM) enzyme that is essential for nitrogenase cofactor assembly. This review focuses on the recent work that sheds light on the role of NifB in the formation of the [Fe8S9C] core of the nitrogenase cofactor, highlighting the structure, function and mechanism of this unique radical SAM methyltransferase.
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- 2022
8. A novel PCR-based genotyping method for Proteus mirabilis – Intergenic region polymorphism analysis
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Kong, Nianqing, Hu, Yilin, Lan, Chenglu, and Bi, Shuilian
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- 2024
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9. Second and Outer Coordination Sphere Effects in Nitrogenase, Hydrogenase, Formate Dehydrogenase, and CO Dehydrogenase.
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Stripp, Sven, Duffus, Benjamin, Fourmond, Vincent, Léger, Christophe, Leimkühler, Silke, Hirota, Shun, Jasniewski, Andrew, Ogata, Hideaki, Hu, Yilin, and Ribbe, Markus
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Aldehyde Oxidoreductases ,Carbon Dioxide ,Formate Dehydrogenases ,Hydrogenase ,Multienzyme Complexes ,Nitrogenase ,Oxidation-Reduction - Abstract
Gases like H2, N2, CO2, and CO are increasingly recognized as critical feedstock in green energy conversion and as sources of nitrogen and carbon for the agricultural and chemical sectors. However, the industrial transformation of N2, CO2, and CO and the production of H2 require significant energy input, which renders processes like steam reforming and the Haber-Bosch reaction economically and environmentally unviable. Nature, on the other hand, performs similar tasks efficiently at ambient temperature and pressure, exploiting gas-processing metalloenzymes (GPMs) that bind low-valent metal cofactors based on iron, nickel, molybdenum, tungsten, and sulfur. Such systems are studied to understand the biocatalytic principles of gas conversion including N2 fixation by nitrogenase and H2 production by hydrogenase as well as CO2 and CO conversion by formate dehydrogenase, carbon monoxide dehydrogenase, and nitrogenase. In this review, we emphasize the importance of the cofactor/protein interface, discussing how second and outer coordination sphere effects determine, modulate, and optimize the catalytic activity of GPMs. These may comprise ionic interactions in the second coordination sphere that shape the electron density distribution across the cofactor, hydrogen bonding changes, and allosteric effects. In the outer coordination sphere, proton transfer and electron transfer are discussed, alongside the role of hydrophobic substrate channels and protein structural changes. Combining the information gained from structural biology, enzyme kinetics, and various spectroscopic techniques, we aim toward a comprehensive understanding of catalysis beyond the first coordination sphere.
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- 2022
10. Characterization of a Nitrogenase Iron Protein Substituted with a Synthetic [Fe4 Se4 ] Cluster.
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Solomon, Joseph B, Tanifuji, Kazuki, Lee, Chi Chung, Jasniewski, Andrew J, Hedman, Britt, Hodgson, Keith O, Hu, Yilin, and Ribbe, Markus W
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Azotobacter vinelandii ,Oxidoreductases ,Nitrogenase ,Iron-Sulfur Proteins ,Oxidation-Reduction ,Biosynthesis ,Catalysis ,Iron Proteins ,[Fe4Se4] Cluster ,Chemical Sciences ,Organic Chemistry - Abstract
The Fe protein of nitrogenase plays multiple roles in substrate reduction and cluster maturation via its redox-active [Fe4 S4 ] cluster. Here we report the synthesis and characterization of a water-soluble [Fe4 Se4 ] cluster that is used to substitute the [Fe4 S4 ] cluster of the Azotobacter vinelandii Fe protein (AvNifH). Biochemical, EPR and XAS/EXAFS analyses demonstrate the ability of the [Fe4 Se4 ] cluster to adopt the super-reduced, all-ferrous state upon its incorporation into AvNifH. Moreover, these studies reveal that the [Fe4 Se4 ] cluster in AvNifH already assumes a partial all-ferrous state ([Fe4 Se4 ]0 ) in the presence of dithionite, where its [Fe4 S4 ] counterpart in AvNifH exists solely in the reduced state ([Fe4 S4 ]1+ ). Such a discrepancy in the redox properties of the AvNifH-associated [Fe4 Se4 ] and [Fe4 S4 ] clusters can be used to distinguish the differential redox requirements for the substrate reduction and cluster maturation of nitrogenase, pointing to the utility of chalcogen-substituted FeS clusters in future mechanistic studies of nitrogenase catalysis and assembly.
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- 2022
11. Evidence of substrate binding and product release via belt-sulfur mobilization of the nitrogenase cofactor
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Lee, Chi Chung, Kang, Wonchull, Jasniewski, Andrew J, Stiebritz, Martin T, Tanifuji, Kazuki, Ribbe, Markus W, and Hu, Yilin
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Inorganic Chemistry ,Chemical Sciences ,Inorganic chemistry ,Physical chemistry ,Chemical engineering - Abstract
The Mo-nitrogenase catalyses the ambient reduction of N2 to NH3 at the M-cluster, a complex cofactor that comprises two metal-sulphur partial cubanes ligated by an interstitial carbide and three belt-sulphurs. A recent crystallographic study suggests binding of N2 via displacement of the belt-sulphur(s) of the M-cluster upon turnover. However, the direct proof of N2 binding and belt-sulphur mobilization during catalysis remains elusive. Here we show that N2 is captured on the M-cluster via electron- and sulphur-depletion, and that the N2-captured state is catalytically competent in generating NH3. Moreover, we demonstrate that product release only occurs when sulphite is supplied along with a reductant, that sulphite is inserted as sulphide into the belt-sulphur displaced positions, and that there is a dynamic in-and-out of the belt-sulphurs during catalysis. Together, these results establish the mobilization of the cofactor belt-sulphurs as a crucial, yet overlooked, mechanistic element of the nitrogenase reaction.
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- 2022
12. Experimental investigation on indoor daylight environment of building with Cadmium Telluride photovoltaic window
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Hu, Yilin, Xue, Qingwen, Wang, Haobo, Zou, Peng, Yang, Jinming, Chen, Shikeng, and Cheng, Yuanda
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- 2024
- Full Text
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13. Visual saliency model based on crowdsourcing eye tracking data and its application in visual design
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Cheng, Shiwei, Fan, Jing, and Hu, Yilin
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- 2023
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14. Decoding emotion with phase–amplitude fusion features of EEG functional connectivity network
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Hu, Liangliang, Tan, Congming, Xu, Jiayang, Qiao, Rui, Hu, Yilin, and Tian, Yin
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- 2024
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15. Integrin-based prognostic model predicts survival, immunotherapy response, and drug sensitivity in gastric cancer
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Hu, Yilin, Chen, Yu, Wu, Menglong, Qian, Chenyu, Chen, Junjie, Wang, Kun, and Xue, Wanjiang
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- 2024
- Full Text
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16. Nitrogenase Fe Protein: A Multi-Tasking Player in Substrate Reduction and Metallocluster Assembly
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Ribbe, Markus W, Górecki, Kamil, Grosch, Mario, Solomon, Joseph B, Quechol, Robert, Liu, Yiling A, Lee, Chi Chung, and Hu, Yilin
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Medicinal and Biomolecular Chemistry ,Organic Chemistry ,Chemical Sciences ,Carbon Dioxide ,Hydrocarbons ,Nitrogenase ,Oxidation-Reduction ,Oxidoreductases ,nitrogenase ,Fe protein ,reductase ,catalysis ,biosynthesis ,FeS enzyme ,C-1 substrate reduction ,C1 substrate reduction ,Theoretical and Computational Chemistry ,Medicinal and biomolecular chemistry ,Organic chemistry - Abstract
The Fe protein of nitrogenase plays multiple roles in substrate reduction and metallocluster assembly. Best known for its function to transfer electrons to its catalytic partner during nitrogenase catalysis, the Fe protein is also a key player in the biosynthesis of the complex metalloclusters of nitrogenase. In addition, it can function as a reductase on its own and affect the ambient reduction of CO2 or CO to hydrocarbons. This review will provide an overview of the properties and functions of the Fe protein, highlighting the relevance of this unique FeS enzyme to areas related to the catalysis, biosynthesis, and applications of the fascinating nitrogenase system.
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- 2022
17. Tracing the incorporation of the “ninth sulfur” into the nitrogenase cofactor precursor with selenite and tellurite
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Tanifuji, Kazuki, Jasniewski, Andrew J, Villarreal, David, Stiebritz, Martin T, Lee, Chi Chung, Wilcoxen, Jarett, Okhi, Yasuhiro, Chatterjee, Ruchira, Bogacz, Isabel, Yano, Junko, Kern, Jan, Hedman, Britt, Hodgson, Keith O, Britt, R David, Hu, Yilin, and Ribbe, Markus W
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Inorganic Chemistry ,Chemical Sciences ,Archaeal Proteins ,Coenzymes ,Density Functional Theory ,Electron Spin Resonance Spectroscopy ,Iron-Sulfur Proteins ,Methanosarcina ,Models ,Chemical ,Nitrogenase ,Selenious Acid ,Sulfur ,Tellurium ,X-Ray Absorption Spectroscopy ,Organic Chemistry ,Chemical sciences - Abstract
Molybdenum nitrogenase catalyses the reduction of N2 to NH3 at its cofactor, an [(R-homocitrate)MoFe7S9C] cluster synthesized via the formation of a [Fe8S9C] L-cluster prior to the insertion of molybdenum and homocitrate. We have previously identified a [Fe8S8C] L*-cluster, which is homologous to the core structure of the L-cluster but lacks the 'ninth sulfur' in the belt region. However, direct evidence and mechanistic details of the L*- to L-cluster conversion upon 'ninth sulfur' insertion remain elusive. Here we trace the 'ninth sulfur' insertion using SeO32- and TeO32- as 'labelled' SO32-. Biochemical, electron paramagnetic resonance and X-ray absorption spectroscopy/extended X-ray absorption fine structure studies suggest a role of the 'ninth sulfur' in cluster transfer during cofactor biosynthesis while revealing the incorporation of Se2-- and Te2--like species into the L-cluster. Density functional theory calculations further point to a plausible mechanism involving in situ reduction of SO32- to S2-, thereby suggesting the utility of this reaction to label the catalytically important belt region for mechanistic investigations of nitrogenase.
- Published
- 2021
18. An EPR and VTVH MCD spectroscopic investigation of the nitrogenase assembly protein NifB.
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Rupnik, Kresimir, Rettberg, Lee, Tanifuji, Kazuki, Rebelein, Johannes G, Ribbe, Markus W, Hu, Yilin, and Hales, Brian J
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S-Adenosylmethionine ,Bacterial Proteins ,Circular Dichroism ,Electron Spin Resonance Spectroscopy ,Protein Conformation ,Protein Binding ,Cofactor ,Electron paramagnetic resonance ,Iron–sulfur cluster ,Magnetic circular dichroism ,Metallocenter assembly ,Nitrogen fixation ,Iron-sulfur cluster ,Inorganic Chemistry ,Medicinal and Biomolecular Chemistry ,Biochemistry and Cell Biology ,Biophysics - Abstract
NifB, a radical SAM enzyme, catalyzes the biosynthesis of the L cluster (Fe8S9C), a structural homolog and precursor to the nitrogenase active-site M cluster ([MoFe7S9C·R-homocitrate]). Sequence analysis shows that NifB contains the CxxCxxxC motif that is typically associated with the radical SAM cluster ([Fe4S4]SAM) involved in the binding of S-adenosylmethionine (SAM). In addition, NifB houses two transient [Fe4S4] clusters (K cluster) that can be fused into an 8Fe L cluster concomitant with the incorporation of an interstitial carbide ion, which is achieved through radical SAM chemistry initiated at the [Fe4S4]SAM cluster upon its interaction with SAM. Here, we report a VTVH MCD/EPR spectroscopic study of the L cluster biosynthesis on NifB, which focuses on the initial interaction of SAM with [Fe4S4]SAM in a variant NifB protein (MaNifBSAM) containing only the [Fe4S4]SAM cluster and no K cluster. Titration of MaNifBSAM with SAM reveals that [Fe4S4]SAM exists in two forms, labeled [Formula: see text] and [Formula: see text]. It is proposed that these forms are involved in the synthesis of the L cluster. Of the two cluster types, only [Formula: see text] initially interacts with SAM, resulting in the generation of Z, an S = ½ paramagnetic [Fe4S4]SAM/SAM complex.
- Published
- 2021
19. Probing the All-Ferrous States of Methanogen Nitrogenase Iron Proteins
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Solomon, Joseph B, Rasekh, Mahtab F, Hiller, Caleb J, Lee, Chi Chung, Tanifuji, Kazuki, Ribbe, Markus W, and Hu, Yilin
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nitrogenase ,Fe protein ,[Fe4S4] cluster ,all-ferrous state ,physiological reduction potential ,CO2 reduction ,hydrocarbon formation ,methanogen - Abstract
The Fe protein of nitrogenase reduces two C1 substrates, CO2 and CO, under ambient conditions when its [Fe4S4] cluster adopts the all-ferrous [Fe4S4]0 state. Here, we show disparate reactivities of the nifH- and vnf-encoded Fe proteins from Methanosarcina acetivorans (designated MaNifH and MaVnfH) toward C1 substrates in the all-ferrous state, with the former capable of reducing both CO2 and CO to hydrocarbons, and the latter only capable of reducing CO to hydrocarbons at substantially reduced yields. EPR experiments conducted at varying solution potentials reveal that MaVnfH adopts the all-ferrous state at a more positive reduction potential than MaNifH, which could account for the weaker reactivity of the MaVnfH toward C1 substrates than MaNifH. More importantly, MaVnfH already displays the g = 16.4 parallel-mode EPR signal that is characteristic of the all-ferrous [Fe4S4]0 cluster at a reduction potential of -0.44 V, and the signal reaches 50% maximum intensity at a reduction potential of -0.59 V, suggesting the possibility of this Fe protein to access the all-ferrous [Fe4S4]0 state under physiological conditions. These results bear significant relevance to the long-lasting debate of whether the Fe protein can utilize the [Fe4S4]0/2+ redox couple to support a two-electron transfer during substrate turnover which, therefore, is crucial for expanding our knowledge of the reaction mechanism of nitrogenase and the cellular energetics of nitrogenase-based processes.
- Published
- 2021
20. X‐Ray Crystallographic Analysis of NifB with a Full Complement of Clusters: Structural Insights into the Radical SAM‐Dependent Carbide Insertion During Nitrogenase Cofactor Assembly
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Kang, Wonchull, Rettberg, Lee A, Stiebritz, Martin T, Jasniewski, Andrew J, Tanifuji, Kazuki, Lee, Chi Chung, Ribbe, Markus W, and Hu, Yilin
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Inorganic Chemistry ,Chemical Sciences ,Underpinning research ,1.1 Normal biological development and functioning ,Crystallography ,X-Ray ,Models ,Molecular ,Molecular Structure ,Nitrogenase ,S-Adenosylmethionine ,carbide insertion ,cofactors ,nitrogenases ,radical SAM enzyme ,structural biology ,Organic Chemistry ,Chemical sciences - Abstract
NifB is an essential radical SAM enzyme required for the assembly of an 8Fe core of the nitrogenase cofactor. Herein, we report the X-ray crystal structures of Methanobacterium thermoautotrophicum NifB without (apo MtNifB) and with (holo MtNifB) a full complement of three [Fe4 S4 ] clusters. Both apo and holo MtNifB contain a partial TIM barrel core, but unlike apo MtNifB, holo MtNifB is fully assembled and competent in cofactor biosynthesis. The radical SAM (RS)-cluster is coordinated by three Cys, and the adjacent K1- and K2-clusters, representing the precursor to an 8Fe cofactor core, are each coordinated by one His and two Cys. Prediction of substrate channels, combined with in silico docking of SAM in holo MtNifB, suggests the binding of SAM between the RS- and K2-clusters and putative paths for entry of SAM and exit of products of SAM cleavage, thereby providing important mechanistic insights into the radical SAM-dependent carbide insertion concomitant with cofactor core formation.
- Published
- 2021
21. Multi-layered electrolytes for solid-state lithium batteries
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Hu, Yilin, Li, Wei, Zhu, Jianxun, Hao, Shu-Meng, Qin, Xuan, Fan, Li-Zhen, Zhang, Liqun, and Zhou, Weidong
- Published
- 2023
- Full Text
- View/download PDF
22. Dishevelled segment polarity protein 2 promotes gastric cancer progression through Wnt/β-catenin pathway
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He, Ruofei, Chen, Yu, Qian, Chenyu, Hu, YiLin, Huang, Xinkun, and Tao, Ran
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- 2023
- Full Text
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23. Interest-driven and legal supervision innovation of the capital pool model in Chinese banking financial products
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Hu, Yilin
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- 2023
- Full Text
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24. Reactivity, Mechanism, and Assembly of the Alternative Nitrogenases
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Jasniewski, Andrew J, Lee, Chi Chung, Ribbe, Markus W, and Hu, Yilin
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Models ,Molecular ,Molybdenum ,Nitrogen ,Nitrogen Fixation ,Nitrogenase ,Chemical Sciences ,General Chemistry - Abstract
Biological nitrogen fixation is catalyzed by the enzyme nitrogenase, which facilitates the cleavage of the relatively inert triple bond of N2. Nitrogenase is most commonly associated with the molybdenum-iron cofactor called FeMoco or the M-cluster, and it has been the subject of extensive structural and spectroscopic characterization over the past 60 years. In the late 1980s and early 1990s, two "alternative nitrogenase" systems were discovered, isolated, and found to incorporate V or Fe in place of Mo. These systems are regulated by separate gene clusters; however, there is a high degree of structural and functional similarity between each nitrogenase. Limited studies with the V- and Fe-nitrogenases initially demonstrated that these enzymes were analogously active as the Mo-nitrogenase, but more recent investigations have found capabilities that are unique to the alternative systems. In this review, we will discuss the reactivity, biosynthetic, and mechanistic proposals for the alternative nitrogenases as well as their electronic and structural properties in comparison to the well-characterized Mo-dependent system. Studies over the past 10 years have been particularly fruitful, though key aspects about V- and Fe-nitrogenases remain unexplored.
- Published
- 2020
25. Structural evidence for a dynamic metallocofactor during N2 reduction by Mo-nitrogenase
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Kang, Wonchull, Lee, Chi Chung, Jasniewski, Andrew J, Ribbe, Markus W, and Hu, Yilin
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Azotobacter vinelandii ,Biocatalysis ,Crystallography ,X-Ray ,Ligands ,Molybdoferredoxin ,Nitrogen ,Oxidation-Reduction ,Protein Multimerization ,Sulfur ,General Science & Technology - Abstract
The enzyme nitrogenase uses a suite of complex metallocofactors to reduce dinitrogen (N2) to ammonia. Mechanistic details of this reaction remain sparse. We report a 1.83-angstrom crystal structure of the nitrogenase molybdenum-iron (MoFe) protein captured under physiological N2 turnover conditions. This structure reveals asymmetric displacements of the cofactor belt sulfurs (S2B or S3A and S5A) with distinct dinitrogen species in the two αβ dimers of the protein. The sulfur-displaced sites are distinct in the ability of protein ligands to donate protons to the bound dinitrogen species, as well as the elongation of either the Mo-O5 (carboxyl) or Mo-O7 (hydroxyl) distance that switches the Mo-homocitrate ligation from bidentate to monodentate. These results highlight the dynamic nature of the cofactor during catalysis and provide evidence for participation of all belt-sulfur sites in this process.
- Published
- 2020
26. Electron Paramagnetic Resonance and Magnetic Circular Dichroism Spectra of the Nitrogenase M Cluster Precursor Suggest Sulfur Migration upon Oxidation: A Proposal for Substrate and Inhibitor Binding.
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Rupnik, Kresimir, Tanifuji, Kazuki, Rettberg, Lee, Ribbe, Markus W, Hu, Yilin, and Hales, Brian J
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Methanosarcina ,Sulfur ,Iron Compounds ,Nitrogenase ,Enzyme Inhibitors ,Circular Dichroism ,Electron Spin Resonance Spectroscopy ,Binding Sites ,Substrate Specificity ,Oxidation-Reduction ,Electrons ,Magnetic Phenomena ,L cluster ,M cluster ,electron paramagnetic resonance ,magnetic circular dichroism ,nitrogenase ,Medicinal and Biomolecular Chemistry ,Biochemistry and Cell Biology ,Organic Chemistry - Abstract
The active site of the nitrogen-fixing enzyme Mo-nitrogenase is the M cluster ([MoFe7 S9 C⋅R-homocitrate]), also known as the FeMo cofactor or FeMoco. The biosynthesis of this highly complex metallocluster involves a series of proteins. Among them, NifB, a radical-SAM enzyme, is instrumental in the assembly of the L cluster ([Fe8 S9 C]), a precursor and all-iron core of the M cluster. In the absence of sulfite, NifB assembles a precursor form of the L cluster called the L* cluster ([Fe8 S8 C]), which lacks the final ninth sulfur. EPR and MCD spectroscopies are used to probe the electronic structures of the paramagnetic, oxidized forms of both the L and L* clusters, labeled LOx and [L*]Ox . This study shows that both LOx and [L*]Ox have nearly identical EPR and MCD spectra, thus suggesting that the two clusters have identical structures upon oxidation; in other words, a sulfur migrates away from LOx following oxidation, thereby rendering the cluster identical to [L*]Ox . It is proposed that a similar migration could occur to the M cluster upon oxidation, and that this is an instrumental part of both M cluster formation and nitrogenase substrate/inhibitor binding.
- Published
- 2020
27. Special Issue on Nitrogenases and Homologous Systems.
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Hu, Yilin and Ribbe, Markus W
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Nitrogenase ,Metalloproteins ,Oxidation-Reduction ,bacteriochlorophyll ,coenzyme F430 ,double-cubane cluster ,nitrogen fixation ,nitrogenase ,Medicinal and Biomolecular Chemistry ,Biochemistry and Cell Biology ,Organic Chemistry - Abstract
The nitrogenase superfamily comprises homologous enzyme systems that carry out fundamentally important processes, including the reduction of N2 and CO, and the biosynthesis of bacteriochlorophyll and coenzyme F430. This special issue provides a cross-disciplinary overview of the ongoing research in this highly diverse and unique research area of metalloprotein biochemistry.
- Published
- 2020
28. Identity and function of an essential nitrogen ligand of the nitrogenase cofactor biosynthesis protein NifB.
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Rettberg, Lee A, Wilcoxen, Jarett, Jasniewski, Andrew J, Lee, Chi Chung, Tanifuji, Kazuki, Hu, Yilin, Britt, R David, and Ribbe, Markus W
- Subjects
Methanosarcina ,Nitrogen ,Nitrogenase ,Alanine ,Histidine ,Bacterial Proteins ,Ligands ,Electron Spin Resonance Spectroscopy ,Mutagenesis ,X-Ray Absorption Spectroscopy - Abstract
NifB is a radical S-adenosyl-L-methionine (SAM) enzyme that is essential for nitrogenase cofactor assembly. Previously, a nitrogen ligand was shown to be involved in coupling a pair of [Fe4S4] clusters (designated K1 and K2) concomitant with carbide insertion into an [Fe8S9C] cofactor core (designated L) on NifB. However, the identity and function of this ligand remain elusive. Here, we use combined mutagenesis and pulse electron paramagnetic resonance analyses to establish histidine-43 of Methanosarcina acetivorans NifB (MaNifB) as the nitrogen ligand for K1. Biochemical and continuous wave electron paramagnetic resonance data demonstrate the inability of MaNifB to serve as a source for cofactor maturation upon substitution of histidine-43 with alanine; whereas x-ray absorption spectroscopy/extended x-ray fine structure experiments further suggest formation of an intermediate that lacks the cofactor core arrangement in this MaNifB variant. These results point to dual functions of histidine-43 in structurally assisting the proper coupling between K1 and K2 and concurrently facilitating carbide formation via deprotonation of the initial carbon radical.
- Published
- 2020
29. Belt-sulfur mobilization in nitrogenase biosynthesis and catalysis
- Author
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Ribbe, Markus W. and Hu, Yilin
- Published
- 2023
- Full Text
- View/download PDF
30. Study on indoor adaptive thermal comfort evaluation method for buildings integrated with semi-transparent photovoltaic window
- Author
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Wang, Haobo, Lin, Chengkai, Hu, Yilin, Zhang, Xingkui, Han, Jun, and Cheng, Yuanda
- Published
- 2023
- Full Text
- View/download PDF
31. Music-emotion EEG coupling effects based on representational similarity
- Author
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Xu, Jiayang, Hu, Liangliang, Qiao, Rui, Hu, Yilin, and Tian, Yin
- Published
- 2023
- Full Text
- View/download PDF
32. Cofactor maturase NifEN: A prototype ancient nitrogenase?
- Author
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Lee, Chi Chung, primary, Górecki, Kamil, additional, Stang, Martin, additional, Ribbe, Markus W., additional, and Hu, Yilin, additional
- Published
- 2024
- Full Text
- View/download PDF
33. Spectroscopic Characterization of an Eight‐Iron Nitrogenase Cofactor Precursor that Lacks the “9th Sulfur”
- Author
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Jasniewski, Andrew J, Wilcoxen, Jarett, Tanifuji, Kazuki, Hedman, Britt, Hodgson, Keith O, Britt, R David, Hu, Yilin, and Ribbe, Markus W
- Subjects
Inorganic Chemistry ,Chemical Sciences ,Coenzymes ,Models ,Molecular ,Molecular Structure ,Nitrogenase ,Spectrum Analysis ,Sulfur ,X-Ray Absorption Spectroscopy ,bioinorganic chemistry ,NifB ,metalloenzymes ,nitrogenase ,iron-sulfur clusters ,Organic Chemistry ,Chemical sciences - Abstract
Nitrogenases catalyze the reduction of N2 to NH4+ at its cofactor site. Designated the M-cluster, this [MoFe7 S9 C(R-homocitrate)] cofactor is synthesized via the transformation of a [Fe4 S4 ] cluster pair into an [Fe8 S9 C] precursor (designated the L-cluster) prior to insertion of Mo and homocitrate. We report the characterization of an eight-iron cofactor precursor (designated the L*-cluster), which is proposed to have the composition [Fe8 S8 C] and lack the "9th sulfur" in the belt region of the L-cluster. Our X-ray absorption and electron spin echo envelope modulation (ESEEM) analyses strongly suggest that the L*-cluster represents a structural homologue to the l-cluster except for the missing belt sulfur. The absence of a belt sulfur from the L*-cluster may prove beneficial for labeling the catalytically important belt region, which could in turn facilitate investigations into the reaction mechanism of nitrogenases.
- Published
- 2019
34. Structural Analysis of a Nitrogenase Iron Protein from Methanosarcina acetivorans: Implications for CO2 Capture by a Surface-Exposed [Fe4S4] Cluster
- Author
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Rettberg, Lee A, Kang, Wonchull, Stiebritz, Martin T, Hiller, Caleb J, Lee, Chi Chung, Liedtke, Jasper, Ribbe, Markus W, Hu, Yilin, Hille, Russ, and Ward, Thomas
- Subjects
Life on Land ,Archaeal Proteins ,Carbon Dioxide ,Crystallization ,Iron ,Iron-Sulfur Proteins ,Methanosarcina ,Nitrogenase ,CO2 capture ,FeS cluster ,iron protein ,methanogen ,nitrogenase ,Microbiology - Abstract
Nitrogenase iron (Fe) proteins reduce CO2 to CO and/or hydrocarbons under ambient conditions. Here, we report a 2.4-Å crystal structure of the Fe protein from Methanosarcina acetivorans (MaNifH), which is generated in the presence of a reductant, dithionite, and an alternative CO2 source, bicarbonate. Structural analysis of this methanogen Fe protein species suggests that CO2 is possibly captured in an unactivated, linear conformation near the [Fe4S4] cluster of MaNifH by a conserved arginine (Arg) pair in a concerted and, possibly, asymmetric manner. Density functional theory calculations and mutational analyses provide further support for the capture of CO2 on MaNifH while suggesting a possible role of Arg in the initial coordination of CO2 via hydrogen bonding and electrostatic interactions. These results provide a useful framework for further mechanistic investigations of CO2 activation by a surface-exposed [Fe4S4] cluster, which may facilitate future development of FeS catalysts for ambient conversion of CO2 into valuable chemical commodities.IMPORTANCE This work reports the crystal structure of a previously uncharacterized Fe protein from a methanogenic organism, which provides important insights into the structural properties of the less-characterized, yet highly interesting archaeal nitrogenase enzymes. Moreover, the structure-derived implications for CO2 capture by a surface-exposed [Fe4S4] cluster point to the possibility of developing novel strategies for CO2 sequestration while providing the initial insights into the unique mechanism of FeS-based CO2 activation.
- Published
- 2019
35. Radical SAM-dependent formation of a nitrogenase cofactor core on NifB
- Author
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Liu, Yiling A., Quechol, Robert, Solomon, Joseph B., Lee, Chi Chung, Ribbe, Markus W., Hu, Yilin, Hedman, Britt, and Hodgson, Keith O.
- Published
- 2022
- Full Text
- View/download PDF
36. Outcomes of laparoscopic versus open total gastrectomy with D2 lymphadenectomy for gastric cancer: a systematic review and meta-analysis
- Author
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Yang, Yongpu, Chen, Yuyan, Hu, Yilin, Feng, Ying, Mao, Qinsheng, and Xue, Wanjiang
- Published
- 2022
- Full Text
- View/download PDF
37. Accurate prediction of dielectric properties and bandgaps in materials with a machine learning approach.
- Author
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Hu, Yilin, Wu, Maokun, Yuan, Miaojia, Wen, Yichen, Ren, Pengpeng, Ye, Sheng, Liu, Fayong, Zhou, Bo, Fang, Hui, Wang, Runsheng, Ji, Zhigang, and Huang, Ru
- Abstract
The conventional approach to exploring suitable dielectrics for future logic and memory devices relies on first-principle calculations, which are expensive and time-consuming. In this work, we adopt a data-driven machine learning (ML)-based approach to build a model for predicting these properties. By incorporating structural information into the input descriptors, we achieve record-high accuracy in predicting the dielectric constant, with the coefficients of determination (R2) of 0.886 and root mean square error (RMSE) of 0.083. Additionally, we achieve high predictions for the bandgap, with accuracies of 0.832 and 0.533 for R2 and RMSE, respectively. The features corresponding to specific properties are analyzed to obtain physical insights. Finally, we employ first-principle calculations to validate the feasibility of this model. This work proposes a highly efficient approach for using ML to predict material properties. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
38. Assembly and Function of Nitrogenase
- Author
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Lee, Chi-Chung, Stiebritz, Martin Tillmann, Hu, Yilin, Ribbe, Markus Walter, Moura, José J. G., editor, Moura, Isabel, editor, and Maia, Luisa B., editor
- Published
- 2021
- Full Text
- View/download PDF
39. Reduction and Condensation of Aldehydes by the Isolated Cofactor of Nitrogenase
- Author
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Lee, Chi Chung, Hu, Yilin, and Ribbe, Markus W
- Subjects
Chemical Sciences - Abstract
Isolated nitrogenase cofactors can reduce CO, CN-, and CO2 to short-chain hydrocarbons in reactions driven by a strong reductant. Here, we use activity analyses and isotope labeling experiments to show that formaldehyde and acetaldehydes can be reduced as-is or reductively condensed into alkanes and alkenes by the isolated cofactor of Mo-nitrogenase in the presence of EuII-diethylenetriamine pentaacetate (DTPA). Further, we demonstrate that aldehydes can be condensed with CO by the isolated cofactor under the same reaction conditions, pointing to aldehyde-derived species as possible intermediates of nitrogenase-catalyzed CO reduction. Our deuterium labeling experiments suggest the formation of a cofactor-bound hydroxymethyl intermediate upon activation of the formaldehyde, as well as the release of C2H4 as a product upon β-hydride elimination of an acetaldehyde-derived hydroxyethyl intermediate. These findings establish the reductive condensation of aldehydes as a previously unobserved reactivity of a biogenic catalyst while at the same time shed light on the mechanism of enzymatic CO reduction and C-C bond formation, thereby providing a useful framework for further exploration of the unique reactivity and potential applications of nitrogenase-based reactions.
- Published
- 2018
40. Probing the coordination and function of Fe4S4 modules in nitrogenase assembly protein NifB.
- Author
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Rettberg, Lee A, Wilcoxen, Jarett, Lee, Chi Chung, Stiebritz, Martin T, Tanifuji, Kazuki, Britt, R David, and Hu, Yilin
- Subjects
Escherichia coli ,Methanosarcina ,Sulfur ,Iron ,Iron Compounds ,Nitrogenase ,S-Adenosylmethionine ,Archaeal Proteins ,Recombinant Fusion Proteins ,Electron Spin Resonance Spectroscopy ,Cloning ,Molecular ,Sequence Alignment ,Gene Expression ,Binding Sites ,Amino Acid Sequence ,Protein Structure ,Secondary ,Protein Binding ,Substrate Specificity ,Genetic Vectors ,Models ,Molecular ,Protein Interaction Domains and Motifs ,Cloning ,Molecular ,Protein Structure ,Secondary ,Models - Abstract
NifB is an essential radical S-adenosylmethionine (SAM) enzyme for nitrogenase cofactor assembly. Previous studies show that NifB couples a putative pair of [Fe4S4] modules (designated K1 and K2) into an [Fe8S9C] cofactor precursor concomitant with radical SAM-dependent carbide insertion through the action of its SAM-binding [Fe4S4] module. However, the coordination and function of the NifB cluster modules remain unknown. Here, we use continuous wave and pulse electron paramagnetic resonance spectroscopy to show that K1- and K2-modules are 3-cysteine-coordinated [Fe4S4] clusters, with a histidine-derived nitrogen serving as the fourth ligand to K1 that is lost upon K1/K2-coupling. Further, we demonstrate that coexistence of SAM/K2-modules is a prerequisite for methyltransfer to K2 and hydrogen abstraction from the K2-associated methyl by a 5'-deoxyadenosyl radical. These results establish an important framework for mechanistic explorations of NifB while highlighting the utility of a synthetic-cluster-based reconstitution approach employed herein in functional analyses of iron-sulfur (FeS) enzymes.
- Published
- 2018
41. Characterization of an M-Cluster-Substituted Nitrogenase VFe Protein
- Author
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Rebelein, Johannes G, Lee, Chi Chung, Newcomb, Megan, Hu, Yilin, Ribbe, Markus W, Buckel, Wolfgang, and Tezcan, Akif
- Subjects
Azotobacter vinelandii ,Carbon Monoxide ,Coenzymes ,Hydrocarbons ,Nitrogenase ,Oxidation-Reduction ,carbon monoxide ,cofactor ,hydrocarbons ,molybdenum ,nitrogenase ,vanadium ,Microbiology - Abstract
The Mo- and V-nitrogenases are two homologous members of the nitrogenase family that are distinguished mainly by the presence of different heterometals (Mo or V) at their respective cofactor sites (M- or V-cluster). However, the V-nitrogenase is ~600-fold more active than its Mo counterpart in reducing CO to hydrocarbons at ambient conditions. Here, we expressed an M-cluster-containing, hybrid V-nitrogenase in Azotobacter vinelandii and compared it to its native, V-cluster-containing counterpart in order to assess the impact of protein scaffold and cofactor species on the differential reactivities of Mo- and V-nitrogenases toward CO. Housed in the VFe protein component of V-nitrogenase, the M-cluster displayed electron paramagnetic resonance (EPR) features similar to those of the V-cluster and demonstrated an ~100-fold increase in hydrocarbon formation activity from CO reduction, suggesting a significant impact of protein environment on the overall CO-reducing activity of nitrogenase. On the other hand, the M-cluster was still ~6-fold less active than the V-cluster in the same protein scaffold, and it retained its inability to form detectable amounts of methane from CO reduction, illustrating a fine-tuning effect of the cofactor properties on this nitrogenase-catalyzed reaction. Together, these results provided important insights into the two major determinants for the enzymatic activity of CO reduction while establishing a useful framework for further elucidation of the essential catalytic elements for the CO reactivity of nitrogenase.IMPORTANCE This is the first report on the in vivo generation and in vitro characterization of an M-cluster-containing V-nitrogenase hybrid. The "normalization" of the protein scaffold to that of the V-nitrogenase permits a direct comparison between the cofactor species of the Mo- and V-nitrogenases (M- and V-clusters) in CO reduction, whereas the discrepancy between the protein scaffolds of the Mo- and V-nitrogenases (MoFe and VFe proteins) housing the same cofactor (M-cluster) allows for an effective assessment of the impact of the protein environment on the CO reactivity of nitrogenase. The results of this study provide a first look into the "weighted" contributions of protein environment and cofactor properties to the overall activity of CO reduction; more importantly, they establish a useful platform for further investigation of the structural elements attributing to the CO-reducing activity of nitrogenase.
- Published
- 2018
42. Tracing the ‘ninth sulfur’ of the nitrogenase cofactor via a semi-synthetic approach
- Author
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Tanifuji, Kazuki, Lee, Chi Chung, Sickerman, Nathaniel S, Tatsumi, Kazuyuki, Ohki, Yasuhiro, Hu, Yilin, and Ribbe, Markus W
- Subjects
Chemical Sciences ,Catalytic Domain ,Iron-Sulfur Proteins ,Models ,Molecular ,Nitrogenase ,S-Adenosylmethionine ,Sulfur ,Organic Chemistry ,Chemical sciences - Abstract
The M-cluster is the [(homocitrate)MoFe7S9C] active site of nitrogenase that is derived from an 8Fe core assembled viacoupling and rearrangement of two [Fe4S4] clusters concomitant with the insertion of an interstitial carbon and a 'ninth sulfur'. Combining synthetic [Fe4S4] clusters with an assembly protein template, here we show that sulfite can give rise to the ninth sulfur that is incorporated in the catalytically important belt region of the cofactor after the radical S-adenosyl-L-methionine-dependent carbide insertion and the concurrent 8Fe-core rearrangement have already taken place. Based on the differential reactivity of the formed cluster species, we also propose a new [Fe8S8C] cluster intermediate, the L*-cluster, which is similar to the [Fe8S9C] L-cluster, but lacks the ninth sulfur from sulfite. This work provides a semi-synthetic tool for protein reconstitution that could be widely applicable for the functional analysis of other FeS systems.
- Published
- 2018
43. A VTVH MCD and EPR Spectroscopic Study of the Maturation of the “Second” Nitrogenase P‑Cluster
- Author
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Rupnik, Kresimir, Lee, Chi Chung, Hu, Yilin, Ribbe, Markus W, and Hales, Brian J
- Subjects
Azotobacter vinelandii ,Bacterial Proteins ,Circular Dichroism ,Electron Spin Resonance Spectroscopy ,Iron-Sulfur Proteins ,Models ,Chemical ,Nitrogenase ,Oxidoreductases ,Inorganic Chemistry ,Physical Chemistry (incl. Structural) ,Other Chemical Sciences ,Inorganic & Nuclear Chemistry - Abstract
The P-cluster of the nitrogenase MoFe protein is a [ Fe8 S7] cluster that mediates efficient transfer of electrons to the active site for substrate reduction. Arguably the most complex homometallic FeS cluster found in nature, the biosynthetic mechanism of the P-cluster is of considerable theoretical and synthetic interest to chemists and biochemists alike. Previous studies have revealed a biphasic assembly mechanism of the two P-clusters in the MoFe protein upon incubation with Fe protein and ATP, in which the first P-cluster is formed through fast fusion of a pair of [ Fe4 S4]+ clusters within 5 min and the second P-cluster is formed through slow fusion of the second pair of [ Fe4 S4]+ clusters in a period of 2 h. Here we report a VTVH MCD and EPR spectroscopic study of the biosynthesis of the slow-forming, second P-cluster within the MoFe protein. Our results show that the first major step in the formation of the second P-cluster is the conversion of one of the precursor [ Fe4 S4]+ clusters into the integer spin cluster [ Fe4 S3-4]α, a process aided by the assembly protein NifZ, whereas the second major biosynthetic step appears to be the formation of a diamagnetic cluster with a possible structure of [ Fe8 S7-8]β, which is eventually converted into the P-cluster.
- Published
- 2018
44. Evaluation of the Catalytic Relevance of the CO‐Bound States of V‐Nitrogenase
- Author
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Lee, Chi Chung, Wilcoxen, Jarett, Hiller, Caleb J, Britt, R David, and Hu, Yilin
- Subjects
Chemical Sciences ,Carbon Monoxide ,Catalysis ,Nitrogen ,Nitrogenase ,Oxidation-Reduction ,Vanadium ,CO binding ,CO reduction ,cofactor ,nitrogenase ,vanadium ,Organic Chemistry ,Chemical sciences - Abstract
Binding and activation of CO by nitrogenase is a topic of interest because CO is isoelectronic to N2 , the physiological substrate of this enzyme. The catalytic relevance of one- and multi-CO-bound states (the lo-CO and hi-CO states) of V-nitrogenase to C-C coupling and N2 reduction was examined. Enzymatic and spectroscopic studies demonstrate that the multiple CO moieties in the hi-CO state cannot be coupled as they are, suggesting that C-C coupling requires further activation and/or reduction of the bound CO entity. Moreover, these studies reveal an interesting correlation between decreased activity of N2 reduction and increased population of the lo-CO state, pointing to the catalytic relevance of the belt Fe atoms that are bridged by the single CO moiety in the lo-CO state. Together, these results provide a useful framework for gaining insights into the nitrogenase-catalyzed reaction via further exploration of the utility of the lo-CO conformation of V-nitrogenase.
- Published
- 2018
45. Identifying ADHD-Related Abnormal Functional Connectivity with a Graph Convolutional Neural Network
- Author
-
Hu, Yilin, primary, Ran, Junling, additional, Qiao, Rui, additional, Xu, Jiayang, additional, Tan, Congming, additional, Hu, Liangliang, additional, and Tian, Yin, additional
- Published
- 2024
- Full Text
- View/download PDF
46. Effect of stability of PTEN on hepatocellular carcinoma
- Author
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Hu, Yilin, primary, Mei, Hongliang, additional, Luan, Tianji, additional, Sun, Huiling, additional, Lu, Qiping, additional, and Liu, Rong, additional
- Published
- 2024
- Full Text
- View/download PDF
47. The in vivo hydrocarbon formation by vanadium nitrogenase follows a secondary metabolic pathway.
- Author
-
Rebelein, Johannes G, Lee, Chi Chung, Hu, Yilin, and Ribbe, Markus W
- Subjects
Azotobacter vinelandii ,Carbon Monoxide ,Hydrocarbons ,Ethane ,Propane ,Ethylenes ,Nitrogenase ,Metabolic Networks and Pathways - Abstract
The vanadium (V)-nitrogenase of Azotobacter vinelandii catalyses the in vitro conversion of carbon monoxide (CO) to hydrocarbons. Here we show that an A. vinelandii strain expressing the V-nitrogenase is capable of in vivo reduction of CO to ethylene (C2H4), ethane (C2H6) and propane (C3H8). Moreover, we demonstrate that CO is not used as a carbon source for cell growth, being instead reduced to hydrocarbons in a secondary metabolic pathway. These findings suggest a possible role of the ancient nitrogenase as an evolutionary link between the carbon and nitrogen cycles on Earth and establish a solid foundation for biotechnological adaptation of a whole-cell approach to recycling carbon wastes into hydrocarbon products. Thus, this study has several repercussions for evolution-, environment- and energy-related areas.
- Published
- 2016
48. ATP‐Independent Turnover of Dinitrogen Intermediates Captured on the Nitrogenase Cofactor.
- Author
-
Lee, Chi Chung, Stang, Martin, Ribbe, Markus W., and Hu, Yilin
- Subjects
NITROGENASES ,NITROGEN ,AZOTOBACTER ,GAS chromatography/Mass spectrometry (GC-MS) ,MOLYBDENUM enzymes ,SPECIES - Abstract
Nitrogenase reduces N2 to NH3 at its active‐site cofactor. Previous studies of an N2‐bound Mo‐nitrogenase from Azotobacter vinelandii suggest binding of three N2 species via asymmetric belt‐sulfur displacements in the two cofactors of its catalytic component (designated Av1*), leading to the proposal of stepwise N2 reduction involving all cofactor belt‐sulfur sites; yet, the evidence for the existence of multiple N2 species on Av1* remains elusive. Here we report a study of ATP‐independent, EuII/SO32−‐driven turnover of Av1* using GC‐MS and frequency‐selective pulse NMR techniques. Our data demonstrate incorporation of D2‐derived D by Av1* into the products of C2H2‐ and H+‐reduction, and decreased formation of NH3 by Av1* concomitant with the release of N2 under H2; moreover, they reveal a strict dependence of these activities on SO32−. These observations point to the presence of distinct N2 species on Av1*, thereby providing strong support for our proposed mechanism of stepwise reduction of N2 via belt‐sulfur mobilization. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
49. Current Understanding of the Biosynthesis of the Unique Nitrogenase Cofactor Core
- Author
-
Hiller, Caleb J., Rettberg, Lee A., Lee, Chi Chung, Stiebritz, Martin T., Hu, Yilin, Mingos, David Michael P., Series Editor, Cardin, Christine, Editorial Board Member, Duan, Xue, Editorial Board Member, Gade, Lutz H., Editorial Board Member, Gómez-Hortigüela Sainz, Luis, Editorial Board Member, Lu, Yi, Editorial Board Member, Macgregor, Stuart A., Editorial Board Member, Neese, Frank, Editorial Board Member, Pariente, Joaquin Perez, Editorial Board Member, Schneider, Sven, Editorial Board Member, Stalke, Dietmar, Editorial Board Member, and Ribbe, Markus W., editor
- Published
- 2019
- Full Text
- View/download PDF
50. Assembly scaffold NifEN: A structural and functional homolog of the nitrogenase catalytic component
- Author
-
Fay, Aaron W, Blank, Michael A, Rebelein, Johannes G, Lee, Chi Chung, Ribbe, Markus W, Hedman, Britt, Hodgson, Keith O, and Hu, Yilin
- Subjects
Generic health relevance ,Azotobacter vinelandii ,Catalytic Domain ,Coenzymes ,Iron ,Iron Chelating Agents ,Molybdenum ,Molybdoferredoxin ,Nitrogenase ,Oxidoreductases ,Protein Binding ,Protein Multimerization ,Protein Subunits ,nitrogenase ,catalysis ,assembly ,functional homolog ,NifEN - Abstract
NifEN is a biosynthetic scaffold for the cofactor of Mo-nitrogenase (designated the M-cluster). Previous studies have revealed the sequence and structural homology between NifEN and NifDK, the catalytic component of nitrogenase. However, direct proof for the functional homology between the two proteins has remained elusive. Here we show that, upon maturation of a cofactor precursor (designated the L-cluster) on NifEN, the cluster species extracted from NifEN is spectroscopically equivalent and functionally interchangeable with the native M-cluster extracted from NifDK. Both extracted clusters display nearly indistinguishable EPR features, X-ray absorption spectroscopy/extended X-ray absorption fine structure (XAS/EXAFS) spectra and reconstitution activities, firmly establishing the M-cluster-bound NifEN (designated NifEN(M)) as the only protein other than NifDK to house the unique nitrogenase cofactor. Iron chelation experiments demonstrate a relocation of the cluster from the surface to its binding site within NifEN(M) upon maturation, which parallels the insertion of M-cluster into an analogous binding site in NifDK, whereas metal analyses suggest an asymmetric conformation of NifEN(M) with an M-cluster in one αβ-half and an empty cluster-binding site in the other αβ-half, which led to the proposal of a stepwise assembly mechanism of the M-cluster in the two αβ-dimers of NifEN. Perhaps most importantly, NifEN(M) displays comparable ATP-independent substrate-reducing profiles to those of NifDK, which establishes the M-cluster-bound αβ-dimer of NifEN(M) as a structural and functional mimic of one catalytic αβ-half of NifDK while suggesting the potential of this protein as a useful tool for further investigations of the mechanistic details of nitrogenase.
- Published
- 2016
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