Crystal structure of the kinase domain of human vascular endothelial growth factor receptor 2: a key enzyme in angiogenesis
Crystal structure of the kinase domain of human vascular endothelial growth factor receptor 2: a key enzyme in angiogenesis
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DOI：
10.1016/s0969-2126(99)80042-2
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发表时间：
1999-03-15
期刊：
影响因子：
--
通讯作者：
Appelt, K
中科院分区：
文献类型：
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作者：
McTigue, MA;Wickersham, JA;Appelt, K
背景：血管生成与肿瘤生长、黄斑变性、视网膜病变等疾病有关。血管内皮生长因子（Vascular endothelial growth factor， VEGF）通过与血管内皮细胞表面的特异性受体（vegfr）结合，刺激血管生成。vegfr是一种受体酪氨酸激酶，与血小板衍生生长因子受体（pdgfr）一样，在激酶结构域中含有一个大的插入物。结果：本文报道了VEGF受体2 （VEGFR2）催化激酶结构域的生成、动力学表征和2.4埃晶体结构。该蛋白结构体缺少68-残基激酶插入结构域（KID）的50个中心残基，与含有整个KID的结构体具有相当的激酶活性。水晶;在无配体磷酸化状态下确定的结构显示出与其他酪氨酸激酶结构相似的整体折叠和催化残基位置。结晶前在Y1059上自磷酸化的激酶激活环大部分是无序的；然而，它的一部分占据抑制底物结合的位置。KID的末端形成一个β样结构，在其他已知的酪氨酸激酶结构中没有观察到，它靠近激酶C末端。结论：大多数VEGFR2 KID残基对激酶活性不是必需的。在KID末端观察到的独特结构也可能出现在其他PDGFR家族成员中，并可能用于正确定向KID进行信号转导。这种VEGFR2激酶结构为设计选择性抗血管生成治疗剂提供了靶点。
Background: Angiogenesis is involved,in tumor growth, macular degeneration, retinopathy and other diseases. Vascular endothelial growth factor (VEGF) stimulates angiogenesis by binding to specific receptors (VEGFRs) on the surface of vascular endothelial cells. VEGFRs are receptor tyrosine kinases that, like the platelet-derived growth factor receptors (PDGFRs), contain a large insert within the kinase domain.Results: We report here the generation, kinetic characterization, and 2.4 Angstrom crystal structure of the catalytic kinase domain of VEGF receptor 2 (VEGFR2). This protein construct, which lacks 50 central residues of the 68-residue kinase insert domain (KID), has comparable kinase activity to constructs containing the entire KID. The crystal; structure, determined in an unliganded phosphorylated state, reveals an overall fold and catalytic residue positions similar to those observed in other tyrosine-kinase structures. The kinase activation loop, autophosphorylated on Y1059 prior to crystallization, is mostly disordered; however, a portion of it occupies a position inhibitory to substrate binding. The ends of the KID form a beta-like structure, not observed in other known tyrosine kinase structures, that packs near to the kinase C terminus.Conclusions: The majority of the VEGFR2 KID residues are not necessary for kinase activity. The unique structure observed for the ends of the KID may also occur in other PDGFR family members and may serve to properly orient the KID for signal transduction. This VEGFR2 kinase structure provides a target for design of selective anti-angiogenic therapeutic agents.