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Mechanism of Kex2p inhibition by its proregion
Author(s) -
Lesage Guillaume,
Tremblay Mélanie,
Guimond Julie,
Boileau Guy
Publication year - 2001
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(01)03096-4
Subject(s) - pichia pastoris , chemistry , recombinant dna , chaperone (clinical) , proteases , biochemistry , microbiology and biotechnology , secretion , enzyme , biology , gene , medicine , pathology
Many proteases are produced as zymogens bearing an N‐terminal proregion acting both as intramolecular chaperone and as enzyme inhibitor. We studied here the inhibition mechanism of the yeast proprotein convertase Kex2p by its proregion. A recombinant secreted and soluble form of Kex2p was produced in Pichia pastoris and its enzymatic properties toward a fluorogenic synthetic peptide were characterized. Recombinant Escherichia coli ‐produced Kex2p proregion specifically and potently inhibited the enzyme, with an IC 50 of 160 nM. Exploration of the inhibition mechanism revealed that the proregion behaved as a mixed inhibitor.