Subcellular localization, partial purification, and characterization of a dynorphin processing endopeptidase from bovine pituitary

Lakshmi Devi, Prem Gupta, Lloyd D. Fricker

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

An enzyme capable of cleaving dynorphin B-29 to dynorphin B-13 is present in bovine pituitary, with 40- to 50-fold higher specific activity in the posterior and intermediate lobes than in the anterior lobe. Subcellular fractionation of bovine neurointermediate pituitary shows that this enzyme is present in the peptide-containing secretory vesicles. The enzyme has been purified 2,800-fold from whole bovine pituitaries using ion-exchange and gel filtration chromatography. Purified dynorphin-converting enzyme has a neutral pH optimum, and is substantially inhibited by the thiol-protease inhibitor p-chloromercuriphenylsulfonic acid, but not by serine or metalloprotease inhibitors. The purified enzyme processes dynorphin B-29 at Arg14, producing both dynorphin B-14 and dynorphin B-13 in a 5:1 ratio. No other cleavages are observed, suggesting that the activity is free from other proteases and is specific for single Arg sequences. Purified enzyme also processes dynorphin A-17 at the single Arg cleavage site, generating both dynorphin A-8 and A-9 in a 7: 1 ratio. The tissue distribution, subcellular localization, and substrate specificity of this enzyme are consistent with a physiological role in the processing of dynorphin B-29 and dynorphin A-17, and possibly other peptides, at single Arg residues.

Original languageEnglish (US)
Pages (from-to)320-329
Number of pages10
JournalJournal of Neurochemistry
Volume56
Issue number1
StatePublished - 1991

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Dynorphins
Endopeptidases
Purification
Enzymes
Processing
Peptides
Ion Exchange
Secretory Vesicles
Metalloproteases
Tissue Distribution
Fractionation
Substrate Specificity
Chromatography
Protease Inhibitors
Sulfhydryl Compounds
Serine
Gel Chromatography
rimorphin
Ion exchange
Peptide Hydrolases

Keywords

  • Endorphin
  • Enkephalin
  • Neuropeptide biosynthesis
  • Opioid peptide
  • Trypsin-like

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Subcellular localization, partial purification, and characterization of a dynorphin processing endopeptidase from bovine pituitary. / Devi, Lakshmi; Gupta, Prem; Fricker, Lloyd D.

In: Journal of Neurochemistry, Vol. 56, No. 1, 1991, p. 320-329.

Research output: Contribution to journalArticle

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