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A rapid fluorometric assay for the proteolytic activity of SKI‐1/S1P based on the surface glycoprotein of the hemorrhagic fever Lassa virus
Author(s) -
Basak Ajoy,
Chrétien Michel,
Seidah Nabil G.
Publication year - 2002
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(02)02394-3
Subject(s) - lassa virus , glycoprotein , chemistry , protease , microbiology and biotechnology , virus , virology , biochemistry , cleavage (geology) , enzyme , biology , paleontology , fracture (geology)
The subtilase subtilisin kexin isozyme‐1 (SKI‐1)/site 1 protease (S1P), has been implicated in the processing of Lassa virus glycoprotein C (GP‐C) precursor into GP1 and GP2 that are responsible for viral fusion with the host cell membrane. Here, we studied in vitro the kinetics of this cleavage by hSKI‐1 using an intramolecularly quenched fluorogenic (IQF) peptide, Q‐GPC 251–263 [Abz‐ 251 Asp‐Ile‐Tyr‐Ile‐Ser‐Arg‐Arg‐Leu‐Leu↓Gly‐Thr‐Phe‐Thr 263 ‐3‐NitroTyr‐Ala‐CONH 2 ], containing the identified site. The measured V max (app) / K m (app) was compared to those for other IQF SKI‐substrates. Q‐GPC 251–263 is cleaved 10‐fold more efficiently than the previously known best SKI‐substrate, Q‐hproSKI 134–142 . This study confirmed the role of SKI‐1 in GP‐C processing and provides a novel, rapid and efficient enzymatic assay of SKI‐1.