Cross-linking of nitrogenase components. Structure and activity of the covalent complex.

A. H. Willing, Millie Georgiadis, D. C. Rees, J. B. Howard

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

The nitrogenase complex from Azotobacter vinelandii is composed of the MoFe protein (Av1), an alpha 2 beta 2 tetramer, and the Fe protein (Av2), a gamma 2 dimer. During turnover of the enzyme, electrons are transferred from Av2 to Av1 in parallel with the hydrolysis of MgATP. Using the cross-linking reagent, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, we have identified some of the properties of the complex between the two components. The cross-linking reaction was highly specific yielding a single apparent Mr = 97,000 protein. The amount of cross-linked product was essentially independent of whether MgATP or MgADP were in the reaction. Also, the amount was maximum at high ratios of Av2 to Av1. The Mr = 97,000 protein was characterized by amino acid analysis and Edman degradation and was found to be consistent with a 1:1 complex of an Av2 gamma subunit and an Av1 beta subunit (the amino terminal serine subunit). The complex was no longer active in the nitrogenase reaction which supports, but does not prove, the requirement for dissociation of the complex after each electron transferred. Nitrogenase activity and cross-linking were inhibited in an identical way by NaCl, which suggests that electrostatic forces are critical to the formation of the electron transfer complex.

Original languageEnglish (US)
Pages (from-to)8499-8503
Number of pages5
JournalJournal of Biological Chemistry
Volume264
Issue number15
StatePublished - May 25 1989
Externally publishedYes

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Nitrogenase
Electrons
Molybdoferredoxin
Adenosine Triphosphate
Ethyldimethylaminopropyl Carbodiimide
Cross-Linking Reagents
Azotobacter vinelandii
Proteins
Electrostatic force
Cross Reactions
Static Electricity
Dimers
Adenosine Diphosphate
Serine
Hydrolysis
Amino Acids
Degradation
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Cross-linking of nitrogenase components. Structure and activity of the covalent complex. / Willing, A. H.; Georgiadis, Millie; Rees, D. C.; Howard, J. B.

In: Journal of Biological Chemistry, Vol. 264, No. 15, 25.05.1989, p. 8499-8503.

Research output: Contribution to journalArticle

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