A sensitive radiometric assay for enkephalin convertase and other carboxypeptidase B-like enzymes

Gary Stack, Lloyd D. Fricker, Solomon H. Snyder

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

A sensitive radiometric assay for carboxypeptidase B-like enzymes has been developed using enkephalin convertase, an enkephalin synthesizing carboxypeptidase. The assay is based on the differential solubility of 3H-labeled substrate and product in chloroform. The substrates 3H-benzoyl-Phe-Ala-Arg or 3H-benzoyl-Phe-Leu-Arg are poorly soluble in chloroform due to the charged arginine. The products of carboxypeptidase B-like activity on these substrates, 3H-benzoyl-Phe-Ala or 3H-benzoyl Phe-Leu partition quantitatively into chloroform, allowing rapid separation of product from substrate. This assay is approximately 100 times more sensitive than a similar fluorometric assay utilizing dansyl-Phe-Ala-Arg as a substrate.

Original languageEnglish (US)
Pages (from-to)113-121
Number of pages9
JournalLife Sciences
Volume34
Issue number2
DOIs
StatePublished - Jan 9 1984
Externally publishedYes

Fingerprint

Carboxypeptidase H
Carboxypeptidase B
Chloroform
Assays
phenylalanylalanine
phenylalanylleucine
Substrates
Enzymes
Solubility
Arginine

ASJC Scopus subject areas

  • Pharmacology

Cite this

A sensitive radiometric assay for enkephalin convertase and other carboxypeptidase B-like enzymes. / Stack, Gary; Fricker, Lloyd D.; Snyder, Solomon H.

In: Life Sciences, Vol. 34, No. 2, 09.01.1984, p. 113-121.

Research output: Contribution to journalArticle

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