Identification of the lysine residue modified during the activation by acetimidylation of horse liver alcohol dehydrogenase
- 28 January 1975
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 14 (2) , 200-203
- https://doi.org/10.1021/bi00673a002
Abstract
A single amino group in horse liver alcohol dehydrogenase was modified with methyl(14C)acetimidate by a differential labeling procedure. Lysine residues outside the active site were modified with ethyl acetimidate while a lysine residue in the active site was protected by the formation of an enzyme-NAD+-pyrazole complex. After the protecting reagents were removed, the enzyme was treated with methyl(14C)acetimidate. Enzyme activity was enhanced 13-fold as 1.1 (14C)acetimidyl group was incorporated per active site. A labeled peptide was isolated from a tryptic-chymotryptic digest of the modified enzyme in 35% overall yield. Amino acid composition and sequential Edman degradations identified the peptide as residues 219-229; lysine residue 228 was modified with the radioactive acetimidyl group.Keywords
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