Kishi J, Hayakawa T
Department of Biochemistry, School of Dentistry, Aichi-Gakuin University, Nagoya, Japan.
Connect Tissue Res. 1989;19(1):63-76. doi: 10.3109/03008208909016815.
Bovine aortic medial tissue and medial smooth muscle cells were demonstrated for the first time to synthesize a latent collagenase together with collagenase inhibitor in culture. Molecular weights of the latent collagenase and its inhibitor derived from aortic medial tissue explant were estimated to be about 52 K by gel filtration and 26.5 K by electrophoresis, respectively. Activated aortic collagenases cleaved type I collagen in solution into 3/4 (alpha A) and 1/4 (alpha B) length cleavage fragments and were inhibited by EDTA, o-phenanthroline, dithiothreitol, bovine serum, and highly purified dental pulp and aortic collagenase inhibitors. The aortic inhibitors showed inhibitory activity against all the animal collagenases tested, except for bacterial collagenase. Double-immunodiffusion analysis using a monospecific antiserum prepared against dental pulp inhibitor showed that the aortic inhibitors are immunologically identical to the pulp inhibitor. Using the same antiserum, we found immunoreactive collagenase inhibitor protein to be localized along the collagen fibers between elastic membranes in aortic medial tissue.
首次证明牛主动脉中膜组织和中膜平滑肌细胞在培养中能合成一种潜在的胶原酶以及胶原酶抑制剂。通过凝胶过滤法估计，源自主动脉中膜组织外植体的潜在胶原酶及其抑制剂的分子量分别约为52K和通过电泳法测得的26.5K。活化的主动脉胶原酶可将溶液中的I型胶原切割成3/4（αA）和1/4（αB）长度的切割片段，并受到乙二胺四乙酸（EDTA）、邻菲罗啉、二硫苏糖醇、牛血清以及高度纯化的牙髓和主动脉胶原酶抑制剂的抑制。主动脉抑制剂对除细菌胶原酶外的所有测试动物胶原酶均显示出抑制活性。使用针对牙髓抑制剂制备的单特异性抗血清进行的双向免疫扩散分析表明，主动脉抑制剂与牙髓抑制剂在免疫上是相同的。使用相同的抗血清，我们发现免疫反应性胶原酶抑制剂蛋白定位于主动脉中膜组织弹性膜之间的胶原纤维上。
