Fulcher Yan G, Sanganna Gari Raghavendar Reddy, Frey Nathan C, Zhang Fuming, Linhardt Robert J, King Gavin M, Van Doren Steven R
Department of Biochemistry and ‡Department of Physics and Astronomy, University of Missouri , Columbia, Missouri 65211, United States.
ACS Chem Biol. 2014 Apr 18;9(4):957-66. doi: 10.1021/cb400898t. Epub 2014 Feb 10.
Activation by glycosaminoglycans (GAGs) is an emerging trend among extracellular proteases important in disease. ProMMP-7, the zymogen of a matrix metalloproteinase secreted by mucosal epithelial and tumor cells, is activated at their surfaces by sulfated GAGs, but how? ProMMP-7 is activated in trans by representative heparin oligosaccharides in a length-dependent manner, with a large jump in activation at lengths of 16 monosaccharides. Imaging by atomic force microscopy visualized small complexes of proMMP-7 molecules linked by 8-mer lengths of heparinoids and extended assembles formed with 16-mer lengths of heparin. Complexes of proMMP-7 with polydisperse heparin or heparan sulfate were more diverse. Heparinoids evidently accelerate activation by tethering multiple proMMP-7 molecules together for proteolytic attack among neighbors. Removal of either the prodomain or C-terminal peptide sequence of KRSNSRKK from MMP-7 prevents formation of the long arrays induced by heparin 16-mers or heparan sulfate. The role of the C-terminus in activation assays suggests it contributes to remote, allosteric binding of GAGs. Enhancement of proteolytic velocity of MMP-by GAGs indicates them to be effectors of V-type allostery. GAGs from proteoglycans appear to assemble proMMP-7 molecules for activation, an event preceding its tumorigenic or antibacterial proteolytic activities at cell surfaces.
由糖胺聚糖(GAGs)激活是在疾病中起重要作用的细胞外蛋白酶的一个新趋势。ProMMP-7是一种由黏膜上皮细胞和肿瘤细胞分泌的基质金属蛋白酶的酶原,它在其表面被硫酸化的GAGs激活,但具体机制如何?ProMMP-7被具有代表性的肝素寡糖以长度依赖的方式反式激活,在16个单糖长度时激活有大幅跃升。通过原子力显微镜成像观察到,ProMMP-7分子的小复合物由8聚体长度的类肝素连接,而16聚体长度的肝素形成延伸的组装体。ProMMP-7与多分散肝素或硫酸乙酰肝素的复合物则更多样化。类肝素显然通过将多个ProMMP-7分子拴在一起,促进相邻分子间的蛋白水解攻击,从而加速激活。去除MMP-7的前结构域或C端肽序列KRSNSRKK,可阻止由16聚体肝素或硫酸乙酰肝素诱导的长阵列形成。C端在激活试验中的作用表明,它有助于GAGs的远程变构结合。GAGs增强MMP的蛋白水解速度,表明它们是V型变构效应物。蛋白聚糖中的GAGs似乎组装ProMMP-7分子以进行激活,这一事件先于其在细胞表面的致瘤或抗菌蛋白水解活性。