Evidence for a protein that enhances the activity of type I procollagen C-proteinase

Abstract

Gel-filtration separated type I procollagen C-proteinase from a glycoprotein that enhanced the enzyme activity by approximately 4-fold. The enhancer was purified by affinity chromatography on a column of Sepharose coupled to the carboxyl propeptide of type I procollagen. Sodium-dodecyl-sulfate- polyacrylamide gel electrophoresis of the affinity-purified enhancer revealed two active major protein bands with molecular weights of 36 and 34 kdal. Both proteins were glycosylated, as shown by binding to concanavalin-A. The enhancer is extremely heat stable (100 degrees C, 15 min) but its activity is totally abolished by treatment with trypsin or bacterial elastase. The enhancer does not alter the digestion intermediates or final products of the enzymatic reaction but it changes the kinetic properties of the reaction, increasing the apparent Km and Vmax values 16- and 20-fold, respectively. It is suggested that the enhancer might play a regulatory role in procollagen processing.