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A shortened synthesis of 4-(3-aminopropyl) pyrazole, an affinity ligand for alcohol dehydrogenase purification
S P Miller1, P R Giri, D Goldman
1Developmental and Metabolic Neurology Branch, National Institutes of Health, Bethesda, MD 20892.
Abstract:
The most efficient, specific and rapid procedures for alcohol dehydrogenase (ADH) purification utilize immobilized 4-(3-aminopropyl) pyrazole to which pyrazole sensitive ADHs, i.e. class I isozymes, bind. Because of the length of the reported synthesis of this affinity resin, we synthesized the 4-(3-aminopropyl) pyrazole ligand by a new method in two steps from commercially available nicotinaldehyde. The ligand synthesized by this simplified procedure was directly coupled to the chain-extended support, Activated CH-Sepharose 4B, to yield the same ligand-spacer combination as reported by L.G. Lange and B.L. Vallee (Biochem. 15: 4681-4686, 1976). Human and hamster class I ADHs purified using this resin were homogeneous by SDS-PAGE followed by silver staining. Specific activity and recovery of human class I ADH were comparable to those previously reported.