Identification and characterization of human endometase (matrix metalloproteinase-26) from endometrial tumor
Identification and characterization of human endometase (matrix metalloproteinase-26) from endometrial tumor
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DOI：
10.1074/jbc.m002349200
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发表时间：
2000-07-07
影响因子：
4.8
通讯作者：
Sang, QXA
中科院分区：
文献类型：
--
作者：
Park, HI;Ni, J;Sang, QXA
我们报告的发现，克隆和一种新的人类基质金属蛋白酶26（MMP-26）（matrixin）基因，子宫内膜酶，子宫内膜肿瘤源性金属蛋白酶的特性。在已测序的300多万个表达序列标签中，子宫内膜酶基因仅从人子宫内膜肿瘤cDNA文库中获得。子宫内膜异位酶mRNA在人子宫中特异性表达，而在检测的其他组织/细胞中不表达，例如睾丸、心脏、脑、肺、肝脏、胸腺和黑色素瘤G361。子宫内膜异位酶蛋白具有信号肽、前肽结构域和催化结构域，其具有独特的“半胱氨酸开关”前肽序列PHCGVPDGSD和锌结合基序VAT-HEIGHSLGLQH。子宫内膜异位酶分别与人金属弹性蛋白酶、基质溶解素、胶原酶-3和基质溶解素具有43%、41%、41%和39%的相同性。酶原的表达和分离，从大肠杆菌的包涵体的分子量为28 kDa。通过蛋白质N-末端测序、银染和免疫印迹分析证实重组蛋白的同一性和均一性。前酶在折叠过程中被部分激活。子宫内膜异位酶选择性切割I型明胶和α（1）-蛋白酶抑制剂;然而，它不消化胶原蛋白、层粘连蛋白、弹性蛋白、β-酪蛋白、纤溶酶原、大豆胰蛋白酶抑制剂或Bowman-Birk抑制剂。它水解基质素和肿瘤坏死因子-α凝聚酶的肽底物。子宫内膜异位酶可以选择性地切割细胞外基质蛋白、丝氨酸蛋白酶抑制剂和加工细胞因子。
We report the discovery, cloning, and characterization of a novel human matrix metalloproteinase 26 (MMP-26) (matrixin) gene, endometase, an endometrial tumor-derived metalloproteinase. Among more than three million expressed sequence tags sequenced, the endometase gene was only obtained from human endometrial tumor cDNA library. Endometase mRNA was expressed specifically in human uterus, not in other tissues/cells tested, e.g. testis, heart, brain, lungs, liver, thymus, and melanoma G361. Endometase protein has a signal peptide, a propeptide domain, and a catalytic domain with a unique "cysteine switch" propeptide sequence, PHCGVPDGSD, and a zinc-binding motif, VAT-HEIGHSLGLQH. Endometase is 43, 41, 41, and 39% identical to human metalloelastase, stromelysin, collagenase-3, and matrilysin, respectively. The zymogen was expressed and isolated from Escherichia coli as inclusion bodies with a molecular mass of 28 kDa. The identity and homogeneity of the recombinant protein was confirmed by protein N-terminal sequencing, silver stain, and immunoblot analyses. The pro-enzyme was partially activated during the folding process. Endometase selectively cleaved type I gelatin and alpha(1)-proteinase inhibitor; however, it did not digest collagens, laminin, elastin, beta-casein, plasminogen, soybean trypsin inhibitor, or Bowman-Birk inhibitor. It hydrolyzed peptide substrates of matrixins and tumor necrosis factor-alpha concerting enzyme. Endometase may selectively cleave extracellular matrix proteins, inactivate serpins, and process cytokines.