Mutagenesis of the proposed iron‐sulfur cluster binding ligands in Escherichia coli biotin synthase

FEBS Letters

Roach, Peter L¹  Shaw, Nicholas M²  Baldwin, Jack E³  Hewitson, Kirsty S³ 

[1] Department of Chemistry, University of Southampton, Highfield, Southampton SO17 1BJ, UK;Biotechnology Department, Lonza AG, CH 3930 Visp, Switzerland;Dyson Perrins Laboratory, University of Oxford, South Parks Road, Oxford OX1 3QY, UK

关键词: Biotin synthase;    Iron-sulfur cluster;    Mutagenesis of iron-sulfur cluster ligand;    SAM;    S-adenosylmethionine;    Fe-S;    iron-sulfur cluster;    DTT;    dithiothreitol;    SDS–PAGE;    sodium dodecyl sulfate–polyacrylamide gel electrophoresis;    pfl AE;    pyruvate formate lyase activating enzyme;    aRR AE;    anaerobic ribonucleotide reductase activating enzyme;   

DOI  :  10.1016/S0014-5793(00)01101-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Biotin synthase (BioB) is a member of a family of enzymes that includes anaerobic ribonucleotide reductase and pyruvate formate lyase activating enzyme. These enzymes all use S-adenosylmethionine during turnover and contain three highly conserved cysteine residues that may act as ligands to an iron-sulfur cluster required for activity. Three mutant enzymes of BioB have been made, each with one cysteine residue (C53, 57, 60) mutated to alanine. All three mutant enzymes were inactive, but they still exhibited the characteristic UV-visible spectrum of a [2Fe-2S]²⁺ cluster similar to that of the wild-type enzyme.

【 授权许可】

Unknown