Contortrostatin, a snake venom disintegrin, inhibits beta 1 integrin-mediated human metastatic melanoma cell adhesion and blocks experimental metastasis.
Contortrostatin, a snake venom disintegrin, inhibits beta 1 integrin-mediated human metastatic melanoma cell adhesion and blocks experimental metastasis.
复制标题
DOI：
--
复制
发表时间：
1994-09
期刊：
影响因子：
11.2
通讯作者：
M. Trikha;Y. Clerck;F. Markland
中科院分区：
文献类型：
--
作者：
M. Trikha;Y. Clerck;F. Markland
去整合素是含有Arg-Gly-Asp的蛋白质，其抑制整合素介导的细胞-细胞和细胞-基质相互作用。我们已经纯化了一个去整合素，contortrostatin，从蝮蛇蛇毒，是一个有效的抑制剂人转移性黑色素瘤（M24 MET）细胞粘附到细胞外基质蛋白。Contortrostatin抑制M24 met细胞与I型胶原、玻连蛋白和纤连蛋白的粘附，50%抑制浓度值分别为20、75和220 nM。Contortrostatin不显著抑制M24 met细胞与层粘连蛋白的粘附。125 I标记的contortrostatin以可饱和和可置换的方式与M24 met细胞结合。Scatchard分析表明，有两个结合位点的125 I标记的contortrostatin在这些细胞的表面上。高亲和力结合具有3 nM的Kd，165，000个位点/细胞，低亲和力结合具有60 nM的Kd，500，000个位点/细胞。固定化的contortrostatin可以支持M24 met细胞的粘附;这种结合被β 1整联蛋白亚基的单克隆抗体和纤连蛋白受体α 5 β 1的抗体阻断。阻断M24 met细胞与固定化玻连蛋白粘附的抗玻连蛋白受体（α v β 5）单克隆抗体不阻断M24 met细胞与固定化contortrostatin的结合。在体内实验转移模型系统中，20 μ g和100 μ g的contortrostatin分别抑制了51%和73%的在scid小鼠尾静脉注射的M24 met细胞（5 × 10（5））的肺定植。我们的结论是，contortrostatin是一种有效的β 1整合素介导的M24细胞粘附在体外的抑制剂，它也抑制肺定植在体内。
Disintegrins are Arg-Gly-Asp-containing proteins that inhibit integrin-mediated cell-cell and cell-matrix interactions. We have purified a disintegrin, contortrostatin, from Agkistrodon contortrix contortrix snake venom that is a potent inhibitor of human metastatic melanoma (M24 met) cell adhesion to extracellular matrix proteins. Contortrostatin inhibits M24 met cell adhesion to type I collagen, vitronectin, and fibronectin with 50% inhibitory concentration values of 20, 75, and 220 nM, respectively. Contortrostatin does not significantly inhibit adhesion of M24 met cells to laminin. 125I-labeled contortrostatin binds to M24 met cells in a saturable and displaceable manner. Scatchard analysis indicates that there are two binding sites for 125I-labeled contortrostatin on the surface of these cells. High affinity binding has a Kd of 3 nM with 165,000 sites/cells low affinity binding has a Kd of 60 nM with 500,000 sites/cell. Immobilized contortrostatin can support adhesion of M24 met cells; this binding is blocked by a monoclonal antibody to the beta 1 integrin subunit and by an antibody to the fibronectin receptor alpha 5 beta 1. The anti-vitronectin receptor (alpha v beta 5) monoclonal antibody which blocks adhesion of M24 met cells to immobilized vitronectin does not block binding of M24 met cells to immobilized contortrostatin. In an in vivo experimental metastasis model system, contortrostatin at 20 micrograms and 100 micrograms inhibits lung colonization of M24 met cells (5 x 10(5)), injected in the tail vein of scid mice, by 51 and 73%, respectively. We conclude that contortrostatin is a potent inhibitor of beta 1 integrin-mediated M24 met cell adhesion in vitro and that it also inhibits lung colonization in vivo.