Primary structure and cDNA cloning of human fibroblast collagenase inhibitor.
Primary structure and cDNA cloning of human fibroblast collagenase inhibitor.
复制标题
人成纤维细胞胶原酶抑制剂的一级结构和cDNA克隆。
DOI：
10.1073/pnas.83.8.2407
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发表时间：
1986
影响因子：
11.1
通讯作者：
G. Stricklin
中科院分区：
文献类型：
--
作者：
D. Carmichael;A. Sommer;R. Thompson;David C. Anderson;Christopher G. Smith;Howard G. WELGUSt;G. Stricklin
我们报道了人成纤维细胞胶原酶抑制物的一级结构和cDNA克隆，这是一种糖蛋白，似乎在调节许多结缔组织来源的金属内切酶的活性方面起着中心作用，包括胶原酶、明胶酶和蛋白聚糖酶。对分泌的人成纤维细胞胶原酶抑制物进行纯化，并进行自动Edman降解。分泌蛋白由184个氨基酸残基组成，含有2个N-连接的寡糖键和6个二硫键。根据该抑制物的特定氨基酸序列合成的寡核苷酸探针用于从人成纤维细胞系中筛选lambda gt10cDNA文库。对两个重叠的克隆进行鉴定，以确定特定mRNA的完整编码和非编码序列。由核苷酸序列推导出的氨基酸序列与蛋白质测序确定的氨基酸序列一致。一个克隆似乎包含完整的5‘端，此外，该cDNA序列预测了一个23个氨基酸的前导肽。另一个克隆代表成熟信息的3‘端，包括一个短的Poly(A)+区。该3‘端序列与已报道的编码小鼠成纤维细胞Poly(A)+RNA蛋白的部分基因非常相似。然而，该抑制物与之前测序的蛋白酶抑制物没有实质性的同源性。
We report the primary structure and cDNA cloning of human fibroblast collagenase inhibitor, a glycoprotein that appears to play a central role in modulating the activity of a number of metalloendoproteases of connective tissue origin including collagenase, gelatinase, and proteoglycanase. Secreted human fibroblast collagenase inhibitor was purified and subjected to automated Edman degradation. The secreted protein consists of 184 amino acid residues; it contains two sites of N-linked oligosaccharide linkage and six disulfide bonds. Synthetic oligonucleotide probes based on selected amino acid sequences of the inhibitor were used to screen a lambda gt10 cDNA library from a human fibroblast line. Two overlapping cDNA clones were characterized to determine the complete coding and noncoding sequences of the specific mRNA. The amino acid sequence deduced from the nucleotide sequence agrees with that determined by protein sequencing. One clone appears to contain the complete 5' end and, in addition, the cDNA sequence predicts a 23-amino acid leader peptide. The other clone represents the 3' end of the mature message and includes a short poly(A)+ tract. This 3' sequence is remarkably similar to a reported cDNA encoding part of the protein derived from mouse fibroblast poly(A)+ RNA. However, this inhibitor has no substantial homology with previously sequenced protease inhibitors.