Purification and partial characterization of a heat-resistant, cytosolic neuropeptidase from rat liver

R. M. Janas, D. L. Marks, Nicholas F La Russo

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

A cholecystokinin octapeptide (CCK-8)-degrading peptidase was purified from rat liver cytosol by heat precipitation of other proteins followed by gel filtration, ion exchange chromatography and preparative gel electrophoresis, using a silicate binding assay to quantitate the degradation of radiolabeled CCK-8. The purified peptidase (M(r) ~ 60,000) had a pH optimum of 6.0; its activity was inhibited by EDTA and 1, 10-phenanthroline but not by phosphoramidon, calpain inhibitor I, bestatin or bacitracin. CCK-8 peptidase rapidly degraded radiolabeled Met-enkephalin as well as 125I-CCK-8, but not a series of other unrelated peptides. Unlabeled Leu-enkephalin, β-casomorphin and neurotensin competitively inhibited the degradation of 125I-CCK-8, suggesting that these opioids are also substrates for the enzyme. These data suggest that this protein is a novel hepatic enzyme which may play a role in the degradation of neuropeptides.

Original languageEnglish (US)
Pages (from-to)574-581
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume198
Issue number2
DOIs
StatePublished - 1994

Fingerprint

Sincalide
Liver
Purification
Rats
Hot Temperature
Degradation
Methionyl Aminopeptidases
Peptide Hydrolases
Gels
Bacitracin
Leucine Enkephalin
Silicates
Neurotensin
Methionine Enkephalin
Ion Exchange Chromatography
Enzymes
Chromatography
Neuropeptides
Electrophoresis
Edetic Acid

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Purification and partial characterization of a heat-resistant, cytosolic neuropeptidase from rat liver. / Janas, R. M.; Marks, D. L.; La Russo, Nicholas F.

In: Biochemical and Biophysical Research Communications, Vol. 198, No. 2, 1994, p. 574-581.

Research output: Contribution to journalArticle

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