Immunohistochemical identification of collagenase in carrageenin granuloma.

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Abstract
The collagenase present in experimental carrageenin granuloma in the guinea pig has been purified to homogeneity in acrylamide gel electrophoresis by a combination of ammonium sulfate salting out and affinity chromatography on Sepharose 4B--collagen-packed columns. The single protein band thus obtained was used as an antigen to obtain a monospecific antibody in heterologous conditions. Several immunodiffusion, immunoaffinity chromatography, and immunoinhibition tests of the antibody against the specific antigen and various possible serum and tissue contaminants suggested that the antibody was specifically directed against the enzyme protein collagenase. Indirect immunohistochemical staining of carrageenin granulomas, samples at different developmental phases with this specific anti-collagenase antibody, revealed that the specific antigenic protein (the enzyme collagenase) is universally present on the extracellular structures at both the collagen-deposition and the collagen-resorption stages. A hypothesis is proposed to account for these findings, namely, that the enzyme collagenase is bound to its substrate (collagen) under both normal and pathological conditions, and that the critical point of control of collagen degradation must be the activation of the collagen-bound enzyme.

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