Action of rheumatoid synovial collagenase on cartilage collagen. Different susceptibilities of cartilage and tendon collagen to collagenase attack.
The action of purified rheumatoid synovial collagenase on purified cartilage collagen, alpha-1(II)-3, in solution at 25 degrees C has been characterised. The enzyme attacked cartilage collagen in solution producing a 58% reduction in specific viscosity and resulting in the appearance of two reaction products which represented approximately three-quarter and one-quarter fragments of the intact molecule as shown by disc electrophoresis in polyacrylamide gels containing sodium dodecyl sulphate. The alpha-chain fragments which comprised each of these components corresponded to molecular weights of approximately 74000 and 21000. Electron microscopy of segment-long-spacing crystallites of the reaction products revealed three-quarter (TC-a) and one-quarter (TC-b) length fragments, and permitted accurate localization of the cleavage locus between bands 41 and 42 (I-41). This cleavage site and the formation of TC-a and TC-b reaction products are very similar to those found for type-I collagen substrates. Cartilage collagen in solution was found to be more resistant to collagenase attack than tendon collagen, the rate of cartilage collagen degradation being six times slower than that for tendon collagen, as judged by viscometry. The mid-point melting temperatures (T-m) for lathyritic cartilage and tendon collagen were 40.5 and 41.5 degrees C, and for the collagenase-produced reaction products 38.5 and 37.5 degrees C, respectively. The significance of these findings is discussed in relation to the structure of type I and II collagens.
D000818:Animals; D001172:Arthritis, Rheumatoid / Q000201:enzymology*; D002356:Cartilage / Q000032:analysis*; D002642:Chick Embryo; D003094:Collagen; D004589:Electrophoresis, Disc; D006801:Humans; D007700:Kinetics; D007842:Lathyrism / Q000378:metabolism; D003012:Microbial Collagenase / Q000378:metabolism*; D008854:Microscopy, Electron; D011489:Protein Denaturation; D013582:Synovial Fluid / Q000201:enzymology*; D013696:Temperature; D013710:Tendons / Q000032:analysis; D013997:Time Factors; D014783:Viscosity
D003094:Collagen; D003012:Microbial Collagenase