title: Direct extraction of gelatinases from rat bone.
abstract: Many studies have shown that gelatinases are secreted into the medium of cultures of various cell and tissue types, including bone cells. It is not clear, however, to what extent the culture process is responsible for inducing the expression of these proteases. In the present study, gelatinolytic enzymes were extracted directly from bone and other tissues and identified as bands of activity on SDS-PAGE enzymograms using gelatin as the substrate. Two forms of gelatinase (72-kDa and 92-kDa) were present in extracts of normal young rat bone. Yields were markedly higher from compact bone than from other tissues (blood, marrow, tendon, cancellous bone, articular cartilage, and skin). More 92-kDa than 72-kDa gelatinase was extracted from bone. The proteolytic specificity of the 92-kDa gelatinase isolated from the bone extract was shown to be similar to that reported for the enzyme isolated from tissue culture media. Native type I collagen was not cleaved but heat denatured type I collagen (gelatin) and native type IV, type V, type IX and type XI collagens were degraded. The proteolytic activity was inhibited by EDTA. The results indicate that more gelatinases can be extracted from bone tissue than from other tissues using mild extraction conditions. The cellular origin and function of these enzymes in bone remain to be defined.
authors.affiliation: Department of Orthopaedics, University of Washington, Seattle.
authors.forename: A M
authors.initials: AM
authors.lastname: Bollen
authors.forename: D R
authors.initials: DR
authors.lastname: Eyre
mesh_terms: D000818:Animals; D001853:Bone Marrow / Q000201:enzymology; D001842:Bone and Bones / Q000201:enzymology*; D002358:Cartilage, Articular / Q000201:enzymology; D003094:Collagen / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005780:Gelatin; D018093:Gelatinases / Q000032:analysis*; D008297:Male; D008970:Molecular Weight; D051381:Rats; D011917:Rats, Inbred Lew; D012867:Skin / Q000201:enzymology; D013710:Tendons / Q000201:enzymology
chemical_list: D005780:Gelatin; D003094:Collagen; D018093:Gelatinases