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

Abstract
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.