Structure-function studies on inhibitory activity of Bungarus multicinctus protease inhibitor-like protein on matrix metalloprotease-2, and invasion and migration of human neuroblastoma SK-N-SH cells
Structure-function studies on inhibitory activity of Bungarus multicinctus protease inhibitor-like protein on matrix metalloprotease-2, and invasion and migration of human neuroblastoma SK-N-SH cells
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DOI：
10.1016/j.toxicon.2009.08.012
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发表时间：
2010-02-01
期刊：
影响因子：
2.8
通讯作者：
Chang, Long-Sen
中科院分区：
文献类型：
--
作者：
Chou, Wen-Min;Liu, Wen-Hsin;Chang, Long-Sen
鉴于几种Kunitz型蛋白抑制剂抑制肿瘤侵袭和转移的发现，本研究的目的是探讨Bungarus multicinctus蛋白酶抑制剂样蛋白2（PILP-2）和PILP-3是否具有抗肿瘤活性。尽管大约 28% 的氨基酸取代发生在 PILP-2 和 PILP-3 之间，但分子模型表明 PILP-2 和 PILP-3 具有相似的折叠结构。与 PILP-2 不同，PILP-3 在消除人神经母细胞瘤 SK-N-SH 细胞的迁移和侵袭方面表现出显着的活性。 PILP-3抑制基质金属蛋白酶2(MMP-2)活性的能力高于PILP-2。 Pull-down 测定揭示了 PILP-3 和 MMP-2 之间的蛋白质-蛋白质相互作用。与N端区域的突变相比，C端氨基酸的替换显着减弱了PILP-3抑制细胞侵袭、细胞迁移和MMP-2活性的能力以及PILP-3与MMP-2的结合能力。分子对接显示PILP-2和PILP-3的N端区域与MMP-2催化结构域活性位点周围的裂口相吻合。与 PILP-2 相比，PILP-3 的 C 端区域被认为与 MMP-2 的催化结构域紧密接触。总的来说，我们的数据表明 PILP-3 是一种 MMP-2 抑制剂，并显示出抑制神经母细胞瘤迁移和侵袭的活性，并表明完整的 C 末端对 PILP-3 的活性至关重要。 (C) 2009 Elsevier Ltd. 保留所有权利。
In view of the findings that several Kunitz-type protein inhibitors suppress tumor invasion and metastasis, the aim of the present study is to explore whether Bungarus multicinctus protease inhibitor-like protein-2 (PILP-2) and PILP-3 exhibit anti-tumor activity. Although approximately 28% of amino acid substitutions occurred between PILP-2 and PILP-3, molecular modeling suggested that PILP-2 and PILP-3 shared similar folded structures. Unlike PILP-2, PILP-3 showed a notable activity in abolishing migration and invasion of human neuroblastoma SK-N-SH cells. The ability of PILP-3 to inhibit matrix metalloprotease-2 (MMP-2) activity was higher than that of PILP-2. Pull-down assay revealed protein-protein interaction between PILP-3 and MMP-2. In contrast to mutation on N-terminal region, replacement of amino acids at C-terminus attenuated notably the ability of PILP-3 to inhibit cell invasion, cell migration and MMP-2 activity as well as the binding capability of PILP-3 with MMP-2. Molecular docking showed that N-terminal region of PILP-2 and PILP-3 fitted into the cleft around the active site of MMP-2 catalytic domain. In contrast to that of PILP-2, C-terminal region of PILP-3 was suggested to be in close contact with catalytic domain of MMP-2. Collectively, our data indicate that PILP-3 is a MMP-2 inhibitor and shows an activity in inhibiting migration and invasion of neuroblastoma, and suggest that intact C-terminus is crucial to the activities of PILP-3. (C) 2009 Elsevier Ltd. All rights reserved.