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PMID: 6326091 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Spectroscopic studies of ferricyanide oxidation of Azotobacter vinelandii ferredoxin I.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 81 ·No. 7 ·1984-04-00 ·Pages 1931-5

Morgan TV, Stephens PJ, Devlin F, Stout CD, Melis KA, Burgess BK

Abstract

The Fe(CN)3-(6) oxidation of the crystallographically characterized [[3Fe-3S], [4Fe-4S]] ferredoxin I of Azotobacter vinelandii has been studied using absorption, circular dichroism, magnetic circular dichroism, and EPR spectroscopies. A paramagnetic intermediate is observed en route to Fe-S cluster-free apoprotein, possessing an anisotropic g approximately equal to 2 EPR signal, surviving to temperatures greater than 77 K. This species is shown to result from 3-electron oxidation of the [4Fe-4S] cluster, without modification of the [3Fe-3S] cluster. However, it does not give rise to observable paramagnetic magnetic circular dichroism in the visible-near UV spectral region and is therefore neither an oxidized HIPIP [4Fe-4S] cluster nor an oxidized [3Fe-3S] cluster. We identify the paramagnetic species as a cysteinyldisulfide radical formed on dissociation of an oxidized cysteinate and an oxidized sulfide ion from the [4Fe-4S] cluster. This conclusion is consistent with the observed reaction stoichiometry, the spectroscopic results obtained, known EPR spectra of disulfide radicals, and the reconstitution of the native [4Fe-4S] cluster by dithiothreitol alone. This reaction, earlier interpreted as a HIPIP-type oxidation, is a previously uncharacterized oxidation reaction of [4Fe-4S] clusters.

MeSH Terms
Azotobacter/metabolism Circular Dichroism Electron Spin Resonance Spectroscopy Ferredoxins/metabolism Ferricyanides Oxidation-Reduction Protein Conformation Spectrophotometry
Chemicals
Ferredoxins Ferricyanides
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Morgan T V
Stephens P J
Devlin F
Stout C D
Melis K A
Burgess B K
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25 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.81.7.1931
Published
1984-04-00
Pages
1931-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC345410
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.81.7.1931 PMC Full Text View journal details
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