Neutral protease cleaving the N-terminal propeptide of type III procollagen: partial purification and characterization of the enzyme from smooth muscle cells of bovine aorta
- 13 March 1984
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 23 (6), 1251-1256
- https://doi.org/10.1021/bi00301a036
Abstract
Procollagen type III amino-terminal protease was detected in cultures of smooth muscle cells of fetal calf aorta, and this protease was purified about 400-fold. Only about half of the enzyme activity was consistently attached to concanavalin A-agarose (Con A-agarose). After affinity chromatography of type III pN-collagen-Sepharose, the Con A bound fraction showed only 1 major band with a MW of about 72,000, this value corresponding well with the elution position of enzyme activity in gel filtration. The enzyme did not cleave procollagens type I or type IV, and denatured type III pN-collagen also remained uncleaved. The Km of the enzyme activity for iodo[14C]acetamide-labeled type III pN-collagen was 0.76 .mu.M. Neutral pH and Ca2+ were required for maximal enzymic activity. The metal chelators ethylenediaminetetraacetic acid and ethylene glycol bis(.beta.-aminoethyl ether)-N,N,N'',N''-tetraacetic acid inhibited the activity as well as did dithiothreitol, but there was only little if any inhibition by several other proteinase inhibitors tested.This publication has 18 references indexed in Scilit:
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