A Diazo Compound for the Photochemical Labeling of Yeast Alcohol Dehydrogenase
1971; Elsevier BV; Volume: 246; Issue: 14 Linguagem: Inglês
10.1016/s0021-9258(18)62036-8
ISSN1083-351X
AutoresDouglas T. Browne, Stephen S. Hixson, F. H. Westheimer,
Tópico(s)Enzyme Catalysis and Immobilization
ResumoAbstract The [14C]diazoacetate ester of 3-hydroxymethylpyridine (I) has been synthesized and exchanged enzymically with DPN+ to form the 3-diazoacetoxymethyl analogue of DPN+ (II). This analogue binds to yeast alcohol dehydrogenase (and to several other dehydrogenases) with association constants comparable to those of the coenzyme; on photolysis, the diazo group is decomposed, with concomitant covalent attachment of radioactive label to the protein. Hydrolysis of the labeled protein, followed by ion exchange chromatography, resulted in the isolation of a number of radioactive materials, among which carboxymethylcysteine has been identified. The limitations of this method of labeling proteins near a particular binding site are discussed.
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