Chemical Science
| Autor: | Casey Van Stappen, Emilio Jiménez-Vicente, Ana Pérez-González, Zhi-Yong Yang, Lance C. Seefeldt, Serena DeBeer, Dennis R. Dean, Laure Decamps |
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| Přispěvatelé: | Royal Society of Chemistry |
| Rok vydání: | 2022 |
| Předmět: | |
| Zdroj: | Chemistry and Biochemistry Faculty Publications |
| ISSN: | 2041-6539 2041-6520 |
| DOI: | 10.1039/d1sc06418e |
| Popis: | Reduction of dinitrogen by molybdenum nitrogenase relies on complex metalloclusters: the [8Fe:7S] P-cluster and the [7Fe:9S:Mo:C:homocitrate] FeMo-cofactor. Although both clusters bear topological similarities and require the reductive fusion of [4Fe:4S] sub-clusters to achieve their respective assemblies, P-clusters are assembled directly on the NifD(2)K(2) polypeptide prior to the insertion of FeMo-co, which is fully assembled separately from NifD(2)K(2). P-cluster maturation involves the iron protein NifH(2) as well as several accessory proteins, whose role has not been elucidated. In the present work, two NifD(2)K(2) species bearing immature P-clusters were isolated from an Azotobacter vinelandii strain in which the genes encoding NifH and the accessory protein NifZ were deleted, and characterized by X-ray absorption spectroscopy and EPR. These analyses showed that both NifD(2)K(2) complexes harbor clusters that are electronically and structurally similar, with each NifDK unit containing two [4Fe:4S](2+/+) clusters. Binding of the accessory protein NifW parallels a decrease in the distance between these clusters, as well as a subtle change in their coordination. These results support a conformational role for NifW in P-cluster biosynthesis, bringing the two [4Fe:4S] precursors closer prior to their fusion, which may be crucial in challenging cellular contexts. Max-Planck Society; DFG SPP 1927 "Iron-Sulfur for Life" [DE 1877/1-1]; Peter und Traudl Engelhorn Stiftung; U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences [DE-AC02-76SF00515]; Bill and Melinda Gates Foundation grant BNF Cereals Phase II [OPP1143172]; Bill and Melinda Gates Foundation grant BNF Cereals Phase III [INV-005889]; U.S. Department of Energy, Office of Science, Basic Energy Science [DE-SC0010834, DE-SC0010867]; Bill & Melinda Gates Foundation [OPP1143172, INV-005889] Published version C. V. S., S. D., and L. D. would like to thank the Max-Planck Society for funding. S. D. and C. V. S. acknowledge the DFG SPP 1927 "Iron-Sulfur for Life" (project DE 1877/1-1) for funding. L. D. thanks the Peter und Traudl Engelhorn Sti.ung for funding. Use of the Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, was supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences under Contract No. DE-AC02-76SF00515. C. V. S., S. D., and L. D. gratefully acknowledge Matthew Latimer for his technical assistance during XAS measurements at beamline 9-3. George E. Cutsail III, Justin H. Henthorn, and Patricia Rodr ' iguez Mac ' ia are also thanked for their assistance in XAS data collection. E. J.-V. and A. P. G. were supported by Bill and Melinda Gates Foundation grants BNF Cereals Phase II (OPP1143172) and BNF Cereals Phase III (INV-005889). Work in the laboratories of L. C. S and D. R. D. is supported by grants from the U.S. Department of Energy, Office of Science, Basic Energy Science, DE-SC0010834 and DE-SC0010867, respectively. This work was supported, in whole or in part, by the Bill & Melinda Gates Foundation (OPP1143172, INV-005889). Under the grant conditions of the Foundation, a Creative Commons Attribution 4.0 Generic License has already been assigned to the Author Accepted Manuscript version that might arise from this submission. |
| Databáze: | OpenAIRE |
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