Serrano Solange M T, Kim Junho, Wang Deyu, Dragulev Bojan, Shannon John D, Mann Henning H, Veit Guido, Wagener Raimund, Koch Manuel, Fox Jay W
Laboratório Especial de Toxinologia Aplicada/Center for Applied Toxinology, Instituto Butantan, 05503-900 Sao Paulo, Brazil.
J Biol Chem. 2006 Dec 29;281(52):39746-56. doi: 10.1074/jbc.M604855200. Epub 2006 Oct 13.
Snake venom metalloproteinases (SVMPs) are members of the Reprolysin family of metalloproteinases to which the ADAM (a disintegrin and metalloproteinase) proteins also belong. The disintegrin-like/cysteine-rich domains of the ADAMs have been implicated in their function. In the case of the SVMPs, we hypothesized that these domains could function to target the metalloproteinases to key extracellular matrix proteins or cell surface proteins. Initially we detected interaction of collagen XIV, a fibril-associated collagen with interrupted triple helices containing von Willebrand factor A (VWA) domains, with the PIII SVMP catrocollastatin. Next we investigated whether other VWA domain-containing matrix proteins could support the binding of PIII SVMPs. Using surface plasmon resonance, the PIII SVMP jararhagin and a recombinant cysteine-rich domain from a PIII SVMP were demonstrated to bind to collagen XIV, collagen XII, and matrilins 1, 3, and 4. Jararhagin was shown to cleave these proteins predominantly at sites localized at or near the VWA domains suggesting that it is the VWA domains to which the PIII SVMPs are binding via their cysteine-rich domain. In light of the fact that these extracellular matrix proteins function to stabilize matrix, targeting the SVMPs to these proteins followed by their specific cleavage could promote the destabilization of extracellular matrix and cell-matrix interactions and in the case of capillaries could contribute to their disruption and hemorrhage. Although there is only limited structural homology shared by the cysteine-rich domains of the PIII SVMPs and the ADAMs our results suggest an analogous function for the cysteine-rich domains in certain members of the expanded ADAM family of proteins to target them to VWA domain-containing proteins.
蛇毒金属蛋白酶（SVMPs）是金属蛋白酶解整合素家族的成员，ADAM（一种解整合素和金属蛋白酶）蛋白也属于该家族。ADAM的解整合素样/富含半胱氨酸结构域与其功能有关。对于SVMPs，我们推测这些结构域可能起到将金属蛋白酶靶向关键细胞外基质蛋白或细胞表面蛋白的作用。最初，我们检测到了胶原XIV（一种与原纤维相关的胶原，其包含中断的三螺旋结构，含有血管性血友病因子A（VWA）结构域）与PIII型SVMP卡托科拉他汀之间的相互作用。接下来，我们研究了其他含VWA结构域的基质蛋白是否能支持PIII型SVMPs的结合。利用表面等离子体共振技术，证明PIII型SVMP贾拉哈金和来自PIII型SVMP的重组富含半胱氨酸结构域能与胶原XIV、胶原XII以及基质金属蛋白酶1、3和4结合。结果表明，贾拉哈金主要在VWA结构域或其附近的位点切割这些蛋白，这表明PIII型SVMPs是通过其富含半胱氨酸结构域与VWA结构域结合的。鉴于这些细胞外基质蛋白起到稳定基质的作用，将SVMPs靶向这些蛋白并随后进行特异性切割可能会促进细胞外基质和细胞 - 基质相互作用的不稳定，对于毛细血管而言，可能会导致其破裂和出血。尽管PIII型SVMPs和ADAMs的富含半胱氨酸结构域仅有有限的结构同源性，但我们的结果表明，在扩展的ADAM蛋白家族的某些成员中，富含半胱氨酸结构域具有类似的功能，可将它们靶向含VWA结构域的蛋白。
