Human skin fibroblast stromelysin: structure, glycosylation, substrate specificity, and differential expression in normal and tumorigenic cells.
Human skin fibroblast stromelysin: structure, glycosylation, substrate specificity, and differential expression in normal and tumorigenic cells.
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人皮肤成纤维细胞溶基质素：结构、糖基化、底物特异性以及正常细胞和致瘤细胞中的差异表达。
DOI：
10.1073/pnas.84.19.6725
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发表时间：
1987
影响因子：
11.1
通讯作者：
Goldberg,GI
中科院分区：
文献类型：
--
作者：
Wilhelm,SM;Collier,IE;Kronberger,A;Eisen,AZ;Marmer,BL;Grant,GA;Bauer,EA;Goldberg,GI
我们已经纯化并测定了人基质分解素的完整一级结构，这是一种具有广泛底物特异性的分泌型金属蛋白酶。人基质分解素是以酶前形式合成的，计算大小为53,977 Da，由17个氨基酸组成的长信号肽。原溶血素以两种形式分泌，NaDodSO4/PAGE的表观分子量分别为60和57 kDa。次要的60 kDa多肽是主要的57 kDa蛋白的糖基化形式，含有N-连接的复杂寡糖。酶原被胰酶激活，导致从酶的氨基末端去除84个氨基酸，产生一个45 kDa的活性酶种。人基质分解素能够降解蛋白多糖、纤维连接蛋白、层粘连蛋白和IV型胶原，但不能降解间质I型胶原。该酶不能激活纯化的人成纤维细胞前胶原酶。对其一级结构的分析表明，基质分解素很可能是人类的类似物，即大鼠转运蛋白，后者是一种癌基因转化诱导的蛋白酶。酶在正常细胞和肿瘤细胞中的表达模式表明，体外培养的人皮肤成纤维细胞结构性地分泌基质分解素(每10(6)个细胞每24小时1-2微克)。转化猴病毒40的人胎肺成纤维细胞、转化ras癌基因的人支气管上皮细胞、纤维肉瘤细胞(HT-1080)和黑色素瘤细胞株(A 2058)不表达该酶，也不能用佛波醇12-肉豆蔻酸酯13-乙酸酯处理这些细胞。我们的数据表明，分泌型金属蛋白酶的表达和可能参与肿瘤发生的原因是转化因子和靶细胞之间的特定相互作用，这种作用在不同的物种中可能有所不同。
We have purified and determined the complete primary structure of human stromelysin, a secreted metalloprotease with a wide range of substrate specificities. Human stromelysin is synthesized in a preproenzyme form with a calculated size of 53,977 Da and a 17-amino acid long signal peptide. Prostromelysin is secreted in two forms, with apparent molecular masses on NaDodSO4/PAGE of 60 and 57 kDa. The minor 60-kDa polypeptide is a glycosylated form of the major 57-kDa protein containing N-linked complex oligosaccharides. Zymogen activation by trypsin results in the removal of 84 amino acids from the amino terminus of the enzyme generating a 45-kDa active enzyme species. Human stromelysin is capable of degrading proteoglycan, fibronectin, laminin, and type IV collagen but not interstitial type I collagen. The enzyme is not capable of activating purified human fibroblast procollagenase. Analysis of its primary structure shows that stromelysin is in all likelihood the human analog of rat transin, which is an oncogene transformation-induced protease. The pattern of enzyme expression in normal and tumorigenic cells revealed that human skin fibroblasts in vitro secrete stromelysin constitutively (1-2 micrograms per 10(6) cells per 24 hr). Human fetal lung fibroblasts transformed with simian virus 40, human bronchial epithelial cells transformed with the ras oncogene, fibrosarcoma cells (HT-1080), and a melanoma cell strain (A 2058), do not express this protease nor can the enzyme be induced in these cells by treatment with phorbol 12-myristate 13-acetate. Our data indicate that the expression and the possible involvement of secreted metalloproteases in tumorigenesis result from a specific interaction between the transforming factor and the target cell, which may vary in different species.