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Extraction of collagenase from human gingiva
Author(s) -
Uitto VeliJuka,
Turto Heikki,
Saxén Leena
Publication year - 1978
Publication title -
journal of periodontal research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.31
H-Index - 83
eISSN - 1600-0765
pISSN - 0022-3484
DOI - 10.1111/j.1600-0765.1978.tb00172.x
Subject(s) - collagenase , chemistry , trypsin , chromatography , extraction (chemistry) , size exclusion chromatography , biochemistry , saline , microbial collagenase , enzyme , biology , endocrinology
Collagenase that cleaves the native collagen molecule into two fragments was extracted from a homogenate of human gingiva obtained from subjects with periodontal disease. Collagenase activity was measured using radioactive type I collagen in solution as substrate and separating the reaction products by SDS acrylamide gel electrophoresis. Extraction of collagenase was performed with a neutral salt solution and the extract was fractionated using either ammonium sulfate or dioxane. Gel filtration was necessary for detection of collagenase activity in the extract. Considerable amounts of collagenase apparrently remained in the insoluble fraction of gingival homogenate bound to collagen fibers as indicated by digestion of gingival collagen during incubation at 36°C. Tissuebound collagenase was relased by heating the insoluble homogenate at 60° with calcium. Collagenase extracted either with saline or by the heat‐treatment was mainly in a latent form and was activated by brief exposure to trypsin.