Identification of lysine‐238 of Escherichia coli biotin carboxylase as an ATP‐binding residue

FEBS Letters

Aramaki, Eiji¹  Tokunaga, Eiko¹  Kondo, Hiroki¹  Kazuta, Yasuaki¹ 

[1] Department of Biochemical Engineering and Science, Kyushu Institute of Technology, Iizuka 820, Japan

关键词: Biotin carboxylase;    Acetyl-coenzyme A carboxylase;    Affinity labeling;    Adenosine diphosphopyridoxal;    Adenosine triphosphate-binding site;    Site-directed mutagenesis;    ACC;    acetyl-CoA carboxylase;    BC;    biotin carboxylase;    CPS;    carbamoyl-phosphate synthetase;    AP2-PL;    adenosine diphosphopyridoxal;    IPTG;    isopropyl-β-d-thiogalactopyranoside;    HEPES;    N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid;   

DOI  :  10.1016/S0014-5793(98)00472-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Escherichia coli biotin carboxylase was affinity labeled with adenosine diphosphopyridoxal to identify its ATP binding site. Lysyl endopeptidase digestion of the modified protein, followed by high performance liquid chromatography separation and amino acid sequencing allowed to identify lysine-238 to be the site of modification. Site-directed mutagenesis of this residue into alanine, arginine or glutamine resulted in mutants with much decreased activity. Lysine-238 seems to interact with the γ-phosphate group of ATP but is not involved in catalysis.

【 授权许可】

Unknown