Open AccessProcollagen N‐Proteinase
Author(s) -
TUDERMAN Leena,
PROCKOP Darwin J.
Publication year - 1982
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1982.tb06716.x
Subject(s) - procollagen peptidase , enzyme , biochemistry , chemistry , electrophoresis , denaturation (fissile materials) , microbiology and biotechnology , gel electrophoresis , biology , nuclear chemistry
Procollagen N‐proteinase was purified about 3700‐fold from chick embryo tendons. Electrophoresis of the protein after iodination and denaturation suggested it was homogeneous. However, the native enzyme could not be examined by gel electrophoresis, and therefore homogeneity of the preparation was not conclusively established. Antibodies to the enzyme completely inhibited activity and gave a single precipitant line by double immuno‐ diffusion. The K m for a native procollagen substrate was 0.3‐0.5 μM. The same protein after denaturation inhibited activity. The enzyme did not cleave type IV procollagen from human fibroblasts or a type IV procollagen from a mouse sarcoma. Ca 2+ was required for maximal enzymic activity. The data suggested a second metal requirement, but this was not identified. Reducing agents and metal chelators inhibited activity, but there was little if any inhibition from several inhibitors of other neutral metalloproteinases.