Action of Rheumatoid Synovial Collagenase on Cartilage Caollagen

Abstract

The action of purified rheumatoid synovial collagenase on purified cartilage collagen, U~( I I )~, in solution at 25 "C has been characterised. The enzyme attacked cartilage collagen in solution producing a 5 8 % 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 a-chain fragments which comprised each of these components corresponded to molecular weights of approximately 74000 and 21 000.Electron microscopy of segment-long-spacing crystallites of the reaction products revealed three-quarter (TCA) and one-quarter (TCB) length fragments, and permitted accurate localization of the cleavage locus between bands41 and 42 (1-41). This cleavage site and the formation of TCA and TCB 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,) for lathyritic cartilage and tendon collagen were 40.5 and 41.5 "C, and for the collagenase-produced reaction products 38.5 and 37.5 "C, respectively. The significance of these findings is discussed in relation to the structure of type I and I1 collagens.At 25 "C rheumatoid synovial collagenase, like other mammalian collagenases, has been shown to attack the collagen molecule in its native triple-helical conformation at a specific locus [l-31 yielding reaction products, commonly known as TCA and TCB, which represent three-quarter and one-quarter length fragments of the collagen molecule respectively. These products have been shown to be more susceptible than the native molecules to thermal denaturation [l]. Such findings were obtained using solutions of mammalian, type I, skin collagen having the molecular composition al(I)2a2, whereas collagen extracted from cartilage containing three identical a chains, and having a molecular composition al(II), [4-61, has been reported to be resistant to collagenases derived from gingival and polymorphonuclear leucocytes [7].Since earlier studies had shown that crude preparations of synovial collagenase were able to digest whole slices of articular cartilage [8,9] and more Enzyme. Collagenase (EC 3.4.24.3); trypsin (EC 3.4.21.4).Eur. J. Biochem. 50 (1975) recently, cartilage collagen has been shown to be attacked by rabbit corneal collagenase [lo] and human skin collagenase [l 13, we have compared the action of purified synovial collagenase on purified cartilage (type 11) and tendon (type I) collagens from lathyritic chicks. MATERIALS AND METHODS Preparation and Purification of Rheumatoid Synovial CollagenaseRheumatoid synovial tissue obtained at synovectomies was cut into fragments approximately 4 mm in greatest dimension and explante...

Mycoplasma-induced BALB/c 3T3 collagenase is a mammalian enzyme

“…This is identical with the substrate-specificity exhibited by rheumatoid synovial collagenase (Woolley et al, 1975a). The inability to degrade basement-membrane collagen type IV appears to be a property common to the vertebrate collagenases, which act on the interstitial collagens, types I, II and III (Liotta et al, 1979;Woolley et al, 1978).…”

The Chemistry and Biology of Collagen

“…Bonds of Gly-Ile or Gly-Leu were also cleaved in collagen type II and type III and the specificity of cleavage was the same for these collagens as it was for type I collagen (Miller et al, 1976a). However, the rate of cleavage of type II collagen was approximately one-fifth to one-sixth that of collagen type I or type HI (McCroskery et al, 1975;Woolley et al, 1975a;Miller et al, 1976a;see Fig. 39).…”

“…In TC B (3), the N ' -N ' terminal junction corresponds to the region between bands 41 and 42 shown in collagen (2). Reproduced with permission fromWoolley et al (1975a).copy in SLS aggregates, the position of cleavage of either type I or type II collagen by vertebrate collagenase can be assigned to the inte rband region between bands 41 and 42, approximately 2250 Â from the amino terminus of the helical part of the molecule (Leibovich and Weiss, 1973; Woolley et al, 1975a; see Fig…”

Tumor invasion and host extracellular matrix

“…Firstly, collagenase is elaborated by various normal and neoplastic cells; whereas its limited production by cells in normal tissues suggests a role in collagen remodeling processes, its more frequent occurrence in malignant tissues suggests an important role in pathological collagen resorption. Studies on the susceptibility of the various genetic types of collagens to mammalian collagenase have shown that type III collagen is cleaved at a rate comparable to that of type I collagen, but at approximately five times the rate measured for type II collagen (104,110,111). Type IV and type V collagens are not susceptible to attack by mammalian collagenase, but are each cleaved by specific neutral metalloproteinases (70,75,83).…”