Escherichia coli contains a protein that is homologous in function and N-terminal sequence to the protein encoded by the nifS gene of Azotobacter vinelandii and that can participate in the synthesis of the Fe-S cluster of dihydroxy-acid dehydratase.
In this paper, I report the purification of a protein from Escherichia coli that is very similar in sequence, molecular weight, and the reactions it can catalyze to the protein encoded by the Azotobacter vinelandii nifS gene. This E. coli protein contains pyridoxal phosphate as a cofactor and catalyzes the removal of sulfur from cysteine to form alanine and S0. When dithiothreitol is present along with cysteine, the S0 formed is reduced to S2-. This protein has a reactive sulfhydryl group that is essential for activity. As isolated, this sulfhydryl group appears to be in a disulfide linkage with the sulfhydryl group from the phosphopantetheine moiety of the acyl carrier protein. The purified E. coli protein can mobilize the sulfur from cysteine and contribute it to the formation of a [4Fe-4S] cluster on the apoprotein of E. coli dihydroxy-acid dehydratase. A mechanism is proposed for the early stages of the synthesis of Fe-S clusters using this protein and sulfur in the S0 oxidation state.
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E. I. du Pont de Nemours & Company, Central Research and Development, Experimental Station, Wilmington, Delaware 19880-0328, USA.
D000409:Alanine / Q000378:metabolism; D000595:Amino Acid Sequence; D016948:Azotobacter vinelandii / Q000235:genetics* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003545:Cysteine / Q000378:metabolism; D004220:Disulfides; D004926:Escherichia coli / Q000378:metabolism*; D005798:Genes, Bacterial; D006193:Haemophilus influenzae / Q000378:metabolism; D006836:Hydro-Lyases / Q000378:metabolism*; D007506:Iron-Sulfur Proteins / Q000096:biosynthesis*; D007700:Kinetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009282:Naphthalenesulfonates / Q000494:pharmacology; D010204:Pantetheine / Q000031:analogs & derivatives / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D011732:Pyridoxal Phosphate / Q000032:analysis / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D001426:Bacterial Proteins; D004220:Disulfides; D007506:Iron-Sulfur Proteins; D046911:Macromolecular Substances; D009282:Naphthalenesulfonates; D010446:Peptide Fragments; D013439:Sulfhydryl Reagents; C080001:nifS protein, Bacteria; D010204:Pantetheine; D011732:Pyridoxal Phosphate; C006421:1,5-I-AEDANS; D006836:Hydro-Lyases; C020052:dihydroxyacid dehydratase; D003545:Cysteine; C003129:4'-phosphopantetheine; D000409:Alanine