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Titre: | The proprotein convertase ski-1/s1p in vitro analysis of lassa virus glycoprotein-derived substrates and ex vivo validation of irreversible peptide inhibitors |
Auteur(s): | Knouch, Nabil |
Date de publication: | 18-aoû-2006 |
Editeur: | university bouira |
Référence bibliographique: | American Society for Biochemistry and Molecular Biology |
Résumé: | Herein we designed, synthesized, tested, and validated fluorogenic methylcoumarinamide (MCA) and chloromethylketonepeptides spanning the Lassa virus GPC cleavage site as substrates and inhibitors for the proprotein convertase SKI-1/S1P. The 7-mer MCA (YISRRLL-MCA) and 8-mer MCA (IYISRRLLMCA) are very efficiently cleaved with respect to both the 6-mer MCA (ISRRLL-MCA) and point mutated fluorogenic analogues, except for the 7-mer mutant Y253F. The importance of the P7 phenylic residue was confirmed by digestions of two 16-mer non-fluorogenic peptidyl substrates that differ by a single point mutation (Y253A). Because NMR analysis of these 16-mer peptides did not reveal significant structural differences at recognition motif RRLL, the P7 Tyr residue is likely important in establishing key interactions within the catalytic pocket of SKI-1. Based on these data, we established through analysis of proATF6 and pro-SREBP-2 cellular processing that decanoylated chloromethylketone 7-mer, 6-mer, and 4-mer peptides containing the core RRLL sequence are irreversible and potent ex vivo SKI-1 inhibitors. Although caution must be exercised in using these inhibitors in in vitro reactions, as they can also inhibit the basic amino acid-specific convertase furin, within cells and when used at concentrations <100 M these inhibitors are relatively specific for inhibition of SKI-1 processing events, as opposed to those performed by furin-like convertases |
URI/URL: | http://dspace.univ-bouira.dz:8080/jspui/handle/123456789/6162 |
Collection(s) : | Articles |
Fichier(s) constituant ce document :
Fichier | Description | Taille | Format | |
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J. Biol. Chem.-2006-Pasquato-23471-81.pdf | 589,75 kB | Adobe PDF | Voir/Ouvrir |
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