Purification of carboxypeptidase A using sepharose 4B-bound 3-phenylpropionate

The activity of carboxypeptidase A [EC 3.4.12.2] was inhibited by 3-phenylpropionate derivatives (p-aminocinnamate, 3-p-aminophenylpropionate and 3-p-acetylaminophenylpropionate), and to investigate its use as a ligand for affinity chromatography 3-p-aminophenylpropionate was directly and indirectly...

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Bibliographic Details
Published in:Journal of biochemistry (Tokyo) 1977-05, Vol.81 (5), p.1285-1291
Main Authors: Oshima, G, Nagasawa, K. (Kitasato Univ., Tokyo (Japan). School of pharmaceutical Sciences)
Format: Article
Language:English
Subjects:
Carboxypeptidases - isolation & purification
Carboxypeptidases - metabolism
Chromatography, Affinity - methods
Kinetics
Pancreas - enzymology
Phenylpropionates
Structure-Activity Relationship
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Summary:The activity of carboxypeptidase A [EC 3.4.12.2] was inhibited by 3-phenylpropionate derivatives (p-aminocinnamate, 3-p-aminophenylpropionate and 3-p-acetylaminophenylpropionate), and to investigate its use as a ligand for affinity chromatography 3-p-aminophenylpropionate was directly and indirectly coupled to Sepharose 4B. Carboxypeptidase A was adsorbed only on 3-p-aminophenylpropionate bound to the gel through p-phenylenediamine as a spacer. Carboxypeptidase A from pancreas was purified by a combination of this affinity adsorbent and ion exchange chromatography. The purified carboxypeptidase A had a homogeneity similar to that of a commercial product, as judged by disc gel electrophoresis. The carboxypeptidase activity of Pronase was slightly retarded on the gel column, but could not be separated from its caseinolytic activity. Angiotensin I-converting enzyme [peptidyl dipeptide hydrolase, EC 3.4.15. 1] obtained from hog kidney cortex was not bound to the gel.
ISSN:0021-924X
1756-2651
1756-2651
DOI:10.1093/oxfordjournals.jbchem.a131581