Proocytocin/neurophysin convertase from bovine neurohypophysis and corpus luteum secretory granules: complete purification, structure-function relationships, and competitive inhibitor

Détails

ID Serval
serval:BIB_F2599178DD79
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Proocytocin/neurophysin convertase from bovine neurohypophysis and corpus luteum secretory granules: complete purification, structure-function relationships, and competitive inhibitor
Périodique
Biochemistry
Auteur(s)
Plevrakis  I., Clamagirand  C., Creminon  C., Brakch  N., Rholam  M., Cohen  P.
ISSN
0006-2960 (Print)
Statut éditorial
Publié
Date de publication
03/1989
Volume
28
Numéro
6
Pages
2705-10
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Mar 21
Résumé
Structure-function relationship studies were conducted on the proocytocin/neurophysin endoprotease previously characterized in both bovine neurohypophyseal and corpus luteum granules, using as a reference substrate a synthetic peptide reproducing the entire (1-20) NH2-terminal domain of the precursor. The [D-Arg12] derivative of proocytocin/neurophysin (1-20) was found to be a good competitive inhibitor of the enzyme (Ki = 30 microM), while the [D-Lys11] derivative was not. This allowed the complete purification of two isoforms of the endoprotease (Mr 58,000 and 52,000, respectively) by affinity chromatography using covalently immobilized [D-Arg12] proocytocin/neurophysin (1-20) as the affinity adsorbent. The use of selectively modified or truncated forms of the reference substrate or of the [D-Arg12] competitive inhibitor of the endoprotease established clearly that this basic pair specific convertase is sensitive to modification of the substrate structure either at the basic residues of the cleavage locus or at amino acids around this site (i.e., Pro7 and Gly9). It is concluded that longer distance interactions between amino acids situated on both the NH2 and COOH sides of the basic doublet Lys11Arg12 may contribute to the stabilization of a preferred substrate conformation allowing recognition by the enzyme subsites.
Mots-clé
Amino Acid Sequence Animals Cattle Chromatography, Affinity Chromatography, Gel Corpus Luteum/*enzymology Cytoplasmic Granules/*enzymology Endopeptidases/*isolation & purification/metabolism Female Kinetics Molecular Sequence Data Molecular Weight Oligopeptides/chemical synthesis Peptides/chemical synthesis Pituitary Gland, Posterior/*enzymology Protease Inhibitors Substrate Specificity
Pubmed
Web of science
Création de la notice
28/01/2008 10:34
Dernière modification de la notice
20/08/2019 16:19
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