López-Escalera R, Pardo A
Departamento de Biologia, Facultad de Ciencias, Universidad Nacional Autónoma de México.
Coll Relat Res. 1987 Sep;7(4):249-57. doi: 10.1016/s0174-173x(87)80031-6.
Guinea pig macrophages were obtained by intraperitoneal injection of carrageenin, and in contrast with peritoneal cells from non-stimulated animals, release in vitro an active collagenase into the culture medium. Short time trypsin incubations and treatment of enzyme preparations with 4-APMA failed to reveal latent enzyme activity. Metallopeptidases specific for gelatin paralleled the presence of collagenase in the media of stimulated cells. Raw medium was fractionated by affinity chromatography with heparin-sepharose, collagenase activity bound to heparin sepharose while a gelatinase with an apparent molecular weight of 54,800 did not. When macrophages were grown in vitro in the presence of albumin, the gelatinase activity showed affinity for heparin and an apparent molecular weight of 113,000.
通过腹腔注射角叉菜胶获得豚鼠巨噬细胞,与未受刺激动物的腹腔细胞相比,其在体外会向培养基中释放一种活性胶原酶。短时间胰蛋白酶孵育以及用4-APMA处理酶制剂均未能揭示潜在的酶活性。对明胶具有特异性的金属肽酶与受刺激细胞培养基中胶原酶的存在情况相似。用肝素-琼脂糖亲和层析法对原始培养基进行分级分离,胶原酶活性与肝素琼脂糖结合,而一种表观分子量为54,800的明胶酶则不结合。当巨噬细胞在白蛋白存在的情况下进行体外培养时,明胶酶活性表现出对肝素的亲和力,且表观分子量为113,000。