Identification of 2Fe-2S cysteine ligands in putidaredoxin.
The iron-sulfur center of putidaredoxin is coordinated by four cysteine sulfhydrals. In order to determine which of the six cysteine residues in the protein coordinate the Fe-S center, we have individually mutated cysteine residues 73, 85 and 86 into serines. Of these mutant proteins, only C85S and C73S express holo-protein as evidence by SDS-PAGE and EPR spectroscopy. This leads us to the conclusion that residues 39,45,48, and 86 are the cysteines that coordinate the iron-sulfur center in putidaredoxin.
Gerber
N C
NC
Department of Biochemistry, University of Illinois, Urbana 61801.
Horiuchi
T
T
Koga
H
H
Sligar
S G
SG
D001483:Base Sequence; D003545:Cysteine; D004252:DNA Mutational Analysis; D004578:Electron Spin Resonance Spectroscopy; D005288:Ferredoxins / Q000235:genetics / Q000378:metabolism*; D007506:Iron-Sulfur Proteins / Q000235:genetics / Q000378:metabolism*; D008024:Ligands; D008667:Metalloproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009841:Oligonucleotides; D011549:Pseudomonas
D005288:Ferredoxins; D007506:Iron-Sulfur Proteins; D008024:Ligands; D008667:Metalloproteins; D009841:Oligonucleotides; C100137:putidaredoxin; D003545:Cysteine