PMID: 38849Aug 15, 1979

Affinity labeling of histidine and lysine residue in the adenosine deaminase substrate binding site

Biochimica Et Biophysica Acta
A LucacchiniC A Rossi


1. Adenosine deaminase was inactivated by 9-(4-bromoacetamidobenzyl)-adenine (I) and 9-(2-bromoacetamidobenzyl)adenine (II), two affinity labels. 2. The stoichiometry of the reaction with reagent II is reported: 1 mol reagent is bound per mol inactive enzyme. Amino acid analysis of the 6 N HCl hydrolyzate of the inactive enzyme identified CM-histidine as the main alkylation product. This is the first evidence of the presence of a histidine in the active site region. 3. The alkylation rate and involved amino acid residues were studied for both reagents I and II, at pH 8 and 5.5. The particular reactivity of a lysine near or in the active site is discussed.

Related Concepts

Affinity Labels
Bos indicus
Molecular Sieve Chromatography
Hydrogen-Ion Concentration
Nucleoside Deaminases
Adenosine Deaminase Inhibitors

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