Activation of the precursor of human stromelysin 2 and its interactions with other matrix metalloproteinases
Activation of the precursor of human stromelysin 2 and its interactions with other matrix metalloproteinases
复制标题
DOI：
10.1046/j.1432-1327.1998.2530067.x
复制
发表时间：
1998-04-01
期刊：
影响因子：
--
通讯作者：
Okada, Y
中科院分区：
文献类型：
--
作者：
Nakamura, H;Fujii, Y;Okada, Y
基质金属蛋白酶(MMPs)被合成为非活性酶原(ProMMPs)，随后被多种因素激活以降解细胞外基质(ECM)。在本研究中，我们研究了基质分解素2(MMP10)对基质金属蛋白酶原的分子间激活机制。从12-O-十四酰佛波醇-13-乙酸酯刺激的OSC-20人口腔鳞癌细胞培养上清液中提纯ProMMP10。最终产物在纯化步骤中被部分激活(约为全部活性的38%)，并含有M-r 56000的原基质金属蛋白酶-10，以及M-r 47000、24000和22000的少量蛋白条带。该酶原被4-氨基苯基汞醋酸酯激活，并被加工成M-r47000和24000的活性形式。Phe82-Ser-Ser-Phe-Pro-Gly是Phe82-Ser-Ser-Phe-Pro-Gly，与基质分解素2完全相同。ProMMP9(孕激素酶B)被基质金属蛋白酶-10激活到其全部活性，并被加工成M-r81000、65000、57000和55000这四个低M-r型，前两者在明胶酶谱上显示出蛋白分解活性。NH2末端序列分析表明，81 000、65 000-和57 000-M-r物种具有相同的Phe88-Gln-Thr-Phe-Glu-Gly序列，表明在65 000-和57 000-M-r形式中，Arg87-Phe88多肽键发生断裂，NH2-末端和COOH-末端均被截断。基质金属蛋白酶-10还能激活原基质金属蛋白酶-7(Promatrilysin)，最高可达全部活性的60%，并产生与4-氨基苯基汞酸汞活化相同的活性物种M-r19000。将前基质金属蛋白酶-2(孕激素酶A)或原基质金属蛋白酶-3与基质金属蛋白酶-10共同孵育，不会导致这些原基质金属蛋白酶的激活。这些结果表明，除了先前报道的组织前胶原酶-1(ProMMP1)和中性粒细胞原胶原酶(ProMMP8)的激活外，MMP10还可激活原MMP9和ProMMP7，提示在各种病理条件下，MMP10可能与其他MMPs共同替代MMP3在ECM降解中的作用。
Matrix metalloproteinases (MMP) art synthesized as inactive zymogens (proMMP) and subsequently activated by many factors to degrade the extracellular matrix (ECM). In the present study, we have examined the intermolecular activation mechanisms of proMMP by MMP-10 (stromelysin 2). ProMMP-10 was purified from the culture media of OSC-20 human oral squamous carcinoma cells stimulated with 12-O-tetradecanoylphorbol 13-acetate. The final products are partially activated (approximate to 38% of the full activity) during the purification steps and contain proMMP-10 of M-r 56 000 with minor protein bands of M-r 47 000, 24 000 and 22 000. The zymogen is activated by 4-aminophenylmercuric acetate and processed to the active forms of M-r 47 000 and 24 000. The NH2-terminal sequence of the 47 000- and 24 000-M-r species is Phe82-Ser-Ser-Phe-Pro-Gly, which is identical to that of stromelysin 2. ProMMP-9 (progelatinase B) is activated by MMP-10 to its full activity and processed to the low-M-r species of M-r 81 000, 65 000, 57 000 and 55 000, the former two of which show proteolytic activity on a gelatin zymography. The NH2 terminal sequence analysis indicates that the 81 000, 65 000- and 57 000-M-r species have the identical sequence of Phe88-Gln-Thr-Phe-Glu-Gly, suggesting the cleavage of the Arg87-Phe88 peptide bond for activation and both NH2-terminal and COOH-terminal truncation in the 65 000- and 57 000-M-r forms. MMP-10 also activates proMMP-7 (promatrilysin) up to about 60% of the full activity and generates the same active species of M-r 19 000 as that obtained by activation with 4-aminophenylmercuric acetate. Incubation of proMMP-2 (progelatinase A) or proMMP-3 with MMP-10 does not result in activation of these proMMP. These results indicate that in addition to the previously reported activation of proMMP-1 (tissue procollagenase) and proMMP-8 (neutrophil procollagenase). MMP-10 can also activate proMMP-9 and proMMP-7, and suggest the possibility that MMP-10 may replace a role of MMP-3 in the ECM degradation in concert with other MMP under various pathological conditions.