Fragmentation of fibronectin by inherent autolytic and matrix metalloproteinase activities.
Fragmentation of fibronectin by inherent autolytic and matrix metalloproteinase activities.
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DOI：
10.1016/j.matbio.2010.09.004
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发表时间：
2011-01
期刊：
影响因子：
6.9
通讯作者：
Xu, Xiaoping
中科院分区：
文献类型：
--
作者：
Steffensen, Bjorn;Chen, Zhihua;Pal, Sanjay;Mikhailova, Margarita;Su, Jianrong;Wang, Yao;Xu, Xiaoping
明胶亲和层析纯化的纤维连接蛋白（FN）不稳定，易碎裂。这种断裂归因于固有的自溶蛋白酶活性以及共纯化的基质金属蛋白酶（MMP）。了解FN蛋白水解发生的机制是很重要的，因为FN片段具有不同于完整FN的生物活性。在排除其他血浆源性蛋白酶的作用后，本实验表明，MMP-2将FN切割成不同片段是与固有的FN活性协同发生的。在56°C下有限热处理30分钟，使FN固有的蛋白酶活性失活，从而急剧减少FN的自溶，其方式与丝氨酸蛋白酶抑制剂的存在相似。热处理没有改变细胞对FN的附着，但显著增加了FN对MMP-2酶切的敏感性。MMP-2的羧基端血红素样结构域（PEX）被证明具有MMP-2与FN相互作用所需的关键外域特性，并且在MMP-2变体中，PEX的缺失显著降低了FN的切割速率。为了验证相互作用的特异性，分离的PEX以浓度依赖的方式与MMP-2竞争FN切割。这些结果进一步阐明了固有的自溶丝氨酸蛋白酶样活性和MMP-2对FN断裂的协同作用，为生物学研究中FN的改性制备和处理提供了理论依据。
Fibronectin (FN) purified by gelatin affinity chromatography is unstable and undergoes fragmentation. The cleavage has been ascribed to inherent autolytic protease activities as well as co-purified matrix metalloproteinases (MMP). Understanding the mechanism by which the proteolysis of FN occurs is important, because the FN fragments have biological activities that differ from those of intact FN. Having excluded contributions of other plasma-derived proteases, the present experiments demonstrated that cleavage of FN by MMP-2 to distinct fragments occurred in synergy with inherent FN activities. Limited heat treatment of FN at 56 °C for 30 min inactivated the inherent protease activities sharply reducing autolysis of FN in a manner similar to that seen in the presence of serine proteinase inhibitors. Heat treatment did not alter cell attachment to FN, but significantly increased the susceptibility of FN to enzymatic cleavage by MMP-2. The carboxyl-terminal hemopexin-like domain (PEX) of MMP-2 was shown to possess critical exodomain properties required for the interactions of MMP-2 with FN, and FN was cleaved at a significantly reduced rate by an MMP-2 variant with deletion of PEX. Verifying the specificity of interactions, isolated PEX competed FN cleavage by MMP-2 in a concentration-dependent manner. These results have further elucidated the synergistic contributions of inherent autolytic serine protease-like activities and MMP-2 to fragmentation of FN and provide the rationale and basis for modified preparation and handling of FN used in biological research.
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影响因子：
7
作者：
Dean, Richard A.;Overall, Christopher M.
通讯作者：
Overall, Christopher M.
影响因子：
6.4
作者：
ENGVALL, E;RUOSLAHTI, E
通讯作者：
RUOSLAHTI, E
影响因子：
64.8
作者：
Lasonder, E;Ishihama, Y;Mann, M
通讯作者：
Mann, M
影响因子：
1.9
作者：
Al-Hazmi, Nadia;Thomas, Gareth J.;Whawell, Simon A.
通讯作者：
Whawell, Simon A.
DOI：
10.1111/j.1432-1033.1990.tb19403.x
发表时间：
1990-11-13
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
BANYAI, L;TREXLER, M;PATTHY, L
通讯作者：
PATTHY, L