Brinckerhoff C E, Ruby P L, Austin S D, Fini M E, White H D
J Clin Invest. 1987 Feb;79(2):542-6. doi: 10.1172/JCI112845.
We used a subclone of a rabbit genomic clone for collagenase that cross-hybridizes with human synovial cell messenger RNA (mRNA) to identify a human collagenase complementary DNA (cDNA) clone. The human cDNA clone is 2.1 kilobases (kb) and selects a mRNA transcript of approximately the same size from primary cultures of rheumatoid synovial cells that produce collagenase, but no mRNA is selected from control (nonproducing) synovial fibroblasts. Restriction enzyme analysis and DNA sequence data indicate that our cDNA clone is full length and that it is identical to that recently described for human skin fibroblast collagenase. The cDNA clone identified a single collagenase gene of approximately 17 kb from blots of human genomic DNA. The identity of human skin and synovial cell collagenase and the ubiquity of this enzyme and of its substrates, the interstitial collagens types I, II, and III, imply that common mechanisms controlling collagenolysis throughout the human body may be operative in both normal and disease states.
我们使用了一种与人类滑膜细胞信使核糖核酸（mRNA）发生交叉杂交的胶原酶兔基因组克隆的亚克隆，来鉴定一种人类胶原酶互补脱氧核糖核酸（cDNA）克隆。该人类cDNA克隆为2.1千碱基（kb），从产生胶原酶的类风湿性滑膜细胞原代培养物中筛选出大小大致相同的mRNA转录本，但从对照（不产生胶原酶的）滑膜成纤维细胞中未筛选到mRNA。限制性内切酶分析和DNA序列数据表明，我们的cDNA克隆是全长的，并且与最近描述的人类皮肤成纤维细胞胶原酶相同。该cDNA克隆从人类基因组DNA印迹中鉴定出一个约17 kb的单一胶原酶基因。人类皮肤和滑膜细胞胶原酶的同一性，以及这种酶及其底物（I型、II型和III型间质胶原）的普遍存在，意味着在正常和疾病状态下，控制人体胶原分解的共同机制可能都在起作用。
