Mechanism of Kex2p inhibition by its proregion.
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.
Lesage
G
G
Département de Biochimie, Université de Montréal, P.O. Box 6128, Succursale Centre-ville, H3C 3J7, Montreal, QC, Canada.
Tremblay
M
M
Guimond
J
J
Boileau
G
G
D000595:Amino Acid Sequence; D004792:Enzyme Precursors / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics; D007700:Kinetics; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics; D043484:Proprotein Convertases; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D029701:Saccharomyces cerevisiae Proteins; D015842:Serine Proteinase Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D013381:Subtilisins / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D013917:Thrombin / Q000494:pharmacology
D004792:Enzyme Precursors; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D015842:Serine Proteinase Inhibitors; D043484:Proprotein Convertases; D013381:Subtilisins; D013917:Thrombin; C058751:KEX2 protein, S cerevisiae