Characterization of PC2, a mammalian Kex2 homologue, following expression of the cDNA in microinjected Xenopus oocytes

Abstract

AbstractA human insulinoma cDNA (PC2) that encodes a protein homologous to the Kex2/subtilisin-like proteinases has recently been described [1990, J. Biol. Chem. 265, 2997–3000]. In order to characterise the associated proteinase activity, mRNA encoding PC2 was synthesised in vitro and micro-injected into Xenopus oocytes. The proteinase activity released into the media from oocytes microinjected with PC2 mRNA was assayed using small peptide fluorogenic substrates. Boc.Gln.Arg.Arg aminomethyl coumarin was hydrolysed in a Ca2+-dependent manner, but substrate analogues bearing a single basic aminoacid were not. The substrate specificity, inhibitor profile, and pH optimum of 5.5 were compatible with an involvement of PC2 in prohormone processing in mammalian cells