Bivalent inhibitor of the N-end rule pathway

Détails

ID Serval
serval:BIB_38CF59810962
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Bivalent inhibitor of the N-end rule pathway
Périodique
Journal of Biological Chemistry
Auteur(s)
Kwon  Y. T., Levy  F., Varshavsky  A.
ISSN
0021-9258 (Print)
Statut éditorial
Publié
Date de publication
06/1999
Volume
274
Numéro
25
Pages
18135-9
Notes
Journal Article
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, U.S. Gov't, P.H.S. --- Old month value: Jun 18
Résumé
The N-end rule relates the in vivo half-life of a protein to the identity of its N-terminal residue. Ubr1p, the recognition (E3) component of the Saccharomyces cerevisiae N-end rule pathway, contains at least two substrate-binding sites. The type 1 site is specific for N-terminal basic residues Arg, Lys, and His. The type 2 site is specific for N-terminal bulky hydrophobic residues Phe, Leu, Trp, Tyr, and Ile. Previous work has shown that dipeptides bearing either type 1 or type 2 N-terminal residues act as weak but specific inhibitors of the N-end rule pathway. We took advantage of the two-site architecture of Ubr1p to explore the feasibility of bivalent N-end rule inhibitors, whose expected higher efficacy would result from higher affinity of the cooperative (bivalent) binding to Ubr1p. The inhibitor comprised mixed tetramers of beta-galactosidase that bore both N-terminal Arg (type 1 residue) and N-terminal Leu (type 2 residue) but that were resistant to proteolysis in vivo. Expression of these constructs in S. cerevisiae inhibited the N-end rule pathway much more strongly than the expression of otherwise identical beta-galactosidase tetramers whose N-terminal residues were exclusively Arg or exclusively Leu. In addition to demonstrating spatial proximity between the type 1 and type 2 substrate-binding sites of Ubr1p, these results provide a route to high affinity inhibitors of the N-end rule pathway.
Mots-clé
Binding Sites Dipeptides/pharmacology Enzyme Inhibitors/pharmacology Escherichia coli/enzymology Fungal Proteins/genetics/metabolism Gene Expression Genes, Reporter Ligands Ligases/metabolism Phenotype Protein Binding Saccharomyces cerevisiae/*enzymology/genetics *Saccharomyces cerevisiae Proteins Ubiquinone/metabolism Ubiquitin-Protein Ligases beta-Galactosidase/genetics/metabolism
Pubmed
Web of science
Open Access
Oui
Création de la notice
28/01/2008 12:17
Dernière modification de la notice
20/08/2019 14:28
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