Characterization of the [3Fe-4S] and [4Fe-4S] clusters in unbound PsaC mutants C14D and C51D. Midpoint potentials of the single [4Fe-4S] clusters are identical to FA and FB in bound PsaC of photosystem I.
In a previous paper we showed that the C51D mutant of PsaC contains a [3Fe-4S] cluster in the FA site and a [4Fe-4S] cluster in the FB site and that the C14D mutant contains an uncharacterized cluster in the FB site and a [4Fe-4S] cluster in the FA site [Zhao, J. D., Li, N., Warren, P. V., Golbeck, J. H., & Bryant, D. A. (1992) Biochemistry 31, 5093-5099]. In this paper we describe the electrochemical and electron spin resonance properties of the recombinant C14D and C51D holoproteins after in vitro reinsertion of the iron-sulfur clusters. Unbound PsaC shows no significant resonances in the oxidized state, but the unbound C14D and C51D mutant proteins show an intense set of resonances at g approximately 2.02 and 1.99 characteristic of an oxidized [3Fe-4S]1+/0 cluster. The Em' values for the [3Fe-4S]1+/0 clusters in C14D (FB*) and C51D (FA*) are -98 mV, and both represent one-electron transfers. After reduction with dithionite at pH 10.0, wild-type PsaC shows a broad set of resonances resulting from the superposition of FA- and FB- characterized by a low-field peak at an apparent g value of 2.051 and a high-field trough at an apparent g value of 1.898. The FB resonances in C51D were slightly narrower, with a low-field peak at an apparent g value of 2.039 and high-field trough at an apparent g value of 1.908.(ABSTRACT TRUNCATED AT 250 WORDS)
Yu
L
L
Department of Biochemistry, University of Nebraska, Lincoln 68583-0718.
Zhao
J
J
Lu
W
W
Bryant
D A
DA
Golbeck
J H
JH
D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D000458:Cyanobacteria / Q000737:chemistry; D004563:Electrochemistry; D004578:Electron Spin Resonance Spectroscopy; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D007506:Iron-Sulfur Proteins / Q000737:chemistry*; D008565:Membrane Proteins; D008872:Microwaves; D016296:Mutagenesis; D010084:Oxidation-Reduction; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry; D045331:Photosystem I Protein Complex; D010957:Plasmids; D013696:Temperature
D001426:Bacterial Proteins; D007506:Iron-Sulfur Proteins; D008565:Membrane Proteins; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; C089909:photosystem I, psaB subunit