Subtilisin Kexin Isozyme-1 (SKI-1): Production, purification, inhibitor design and biochemical applications
Subtilisin Kexin Isozyme-1 (SKI-1) belongs to pyrolysin branch of mammalian subtilases [1, 2]. It is synthesized as a membrane bound inactive precursor protein and undergoes auto-catalytic shedding at KHQKLL935⇓ to produce active 98kDa soluble form . This endoprotease cleaves precursor proteins at the motif (R-X-L/I-L/T/K↓), X = any amino acid except Cys. Presence of an additional P7-Tyr greatly enhances SKI-1 recognition . SKI-1 plays roles in cholesterol/lipid synthesis, cartilage development and hemorrhagic fever infections caused by Lassa, Crimean-Congo and Lymphocytic Choriomeningitis viruses . For this reason attention has been focussed on the production of active soluble form of this enzyme and its inhibitors. We have previously described C-terminal modified and pseudopeptide based inhibitors as well as assay of this enzyme . In this study we produced a soluble recombinant human (h) SKI-1 and designed a new miniserpin inhibitor of SKI-1 using the...
KeywordsLymphocytic Choriomeningitis Fever Infection Lassa Virus Enzyme Kinetic Study Tetrapeptide Sequence
Funds from NSERC, Canada.