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PMID: 3006060 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

In vitro synthesis of the iron-molybdenum cofactor of nitrogenase.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 6 ·1986-03-00 ·Pages 1636-40

Shah VK, Imperial J, Ugalde RA, Ludden PW, Brill WJ

Abstract

Molybdate- and ATP-dependent in vitro synthesis of the iron-molybdenum cofactor (FeMo-co) of nitrogenase requires the protein products of at least the nifB, nifN, and nifE genes. Extracts of FeMo-co-negative mutants of Klebsiella pneumoniae and Azotobacter vinelandii with lesions in different genes can be complemented for FeMo-co synthesis. Both K. pneumoniae and A. vinelandii dinitrogenase (component I) deficient in FeMo-co can be activated by FeMo-co synthesized in vitro. Properties of the partially purified dinitrogenase activated by FeMo-co synthesized in vitro were comparable to those of dinitrogenase from the wild-type organism; e.g., ratios of acetylene- to nitrogen-reduction activities, as well as those of acetylene reduction activities to EPR spectrum peak height at g = 3.65, were very similar. A. vinelandii mutants UW45 and CA30 have mutations in a gene functionally equivalent to nifB of K. pneumoniae.

MeSH Terms
Acetylene/metabolism Adenosine Triphosphate/metabolism Azotobacter/enzymology,genetics Bacterial Proteins/genetics,metabolism Cell-Free System Chloramphenicol/pharmacology Electron Spin Resonance Spectroscopy Enzyme Activation Ferredoxins/biosynthesis Genes, Bacterial Genetic Complementation Test Klebsiella pneumoniae/enzymology,genetics Molybdenum/antagonists & inhibitors,pharmacology Molybdoferredoxin/biosynthesis,genetics Nitrogen Fixation Nitrogenase/genetics,metabolism Operon Protein Processing, Post-Translational Tungsten/pharmacology Tungsten Compounds Vanadates Vanadium/pharmacology
Chemicals
Bacterial Proteins Ferredoxins Molybdoferredoxin Tungsten Compounds Vanadium molybdate Vanadates Chloramphenicol Molybdenum Adenosine Triphosphate tetrathiomolybdate Nitrogenase Acetylene tungstate Tungsten
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Shah V K
Imperial J
Ugalde R A
Ludden P W
Brill W J
References (34)
34 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.83.6.1636
Published
1986-03-00
Pages
1636-40
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC323138
Subset
IM
Grants
NIGMS NIH HHS · GM22130 · United States
NCRR NIH HHS · RR-01008 · United States
External Links
PubMed source DOI: 10.1073/pnas.83.6.1636 PMC Full Text View journal details
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