Affinity labeling of the pyridoxal phosphate binding site of the beta2 subunit of Escherichia coli tryptophan synthase.
作者： W Higgins , E W Miles
DOI: 10.1016/S0021-9258(17)30438-6
关键词: Chemistry 、 Pyridoxal phosphate binding 、 Affinity label 、 Protein subunit 、 Tryptophan synthase 、 Affinity labeling 、 Pyridoxal phosphate 、 Cysteine 、 Biochemistry 、 Phosphate
摘要: We have synthesized bromoacetylpyridoxamine phosphate and shown that they meet three criteria for affinity labels of the beta2 subunit tryptophan synthase: (i) kinetic data inactivation indicate a binary complex is formed prior to covalent attachment; (ii) largely prevented by presence pyridoxal phosphate; (iii) stoichiometric with incorporation 0.7 0.8 mol chromophore/mol beta monomer. Our conclusion apo due modification cysteine based on disappearance 1 -SH/beta monomer finding [14C]carboxymethyl derivative in acid hydrolysate protein modified bromo[14C]acetylpyridixamine phosphate. A 39-residue tryptic peptide containing this essential has been isolated purified from bromo[14C]acetylpyridoxamine phosphate-labeled subunit.
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