| Popis: |
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. |
