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 language | English (US) |
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Pages (from-to) | 113-121 |
Number of pages | 9 |
Journal | Life Sciences |
Volume | 34 |
Issue number | 2 |
DOIs | |
State | Published - Jan 9 1984 |
Externally published | Yes |
ASJC Scopus subject areas
- General Pharmacology, Toxicology and Pharmaceutics
- General Biochemistry, Genetics and Molecular Biology