Protein-protein interaction in intracellular signal transduction

细胞内信号转导中的蛋白质-蛋白质相互作用

• 批准号: 10179104
• 负责人: INAGAKI Fuyuhiko
• 金额: $ 84.35万
• 依托单位: Hokkaido University (1999-2001)Tokyo Metropolitan Organization for Medical Research (1998)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas (A)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10179104/
• 关键词: Vav; SH3; Grb2; praline rich region; tertiary structure IP3; radixin; IP3; PTB様ドメイン; Vav-nSH3; PRR; Grb2-cSH3; バキュロウィルス; 多核体病ウィルス; Rho; N-WASP; NMR; X線結晶構造解析; タイトジャンクション; 好中級活性酸素発生系; PBI; PB2; Neurop

Vav is a guanine nucleotide exchange factor for the Rho/Rac family that is expressed exclusively in hematopoietic cells. Growth factor receptor-bound protein 2 (Grb2) has been proposed to play important roles in the membrane localization and activation of Vav through dimerization of its C-terminal Srchomology 3 (SH3) domain (GrbS) and the N-terminal SH3 domain of Vav (VavS). The crystal strucuture of VavS complexed with GrbS has been solved. VavS is distinct from other SH3 domain proteins in that its binding site for proline-rich peptides is blocked by its own RT loop. One of the ends of the VavS β-barrel forms a concave hydrophobic surface. The GrbS components make a contiguous complementary interface with the VavS surface. The binding site of GrbS for VavS partially overlaps with the canonical binding site for proline-rich peptides, but is definitely different. Mutations at the interface caused a decrease in the binding affinity of VavS for GrbS by 4- to 40-fold. The structure reveals how GrbS discriminates VavS specifically from other signaling molecules without binding to the proline-rich motif.

Vav是Rho/Rac家族的鸟嘌呤核苷酸交换因子，仅在造血细胞中表达。生长因子受体结合蛋白2（Growth factor receptor binding protein 2，Grb 2）通过其C端S端结构域（Srchomology 3 domain，Grb S）和N端S端结构域（Vav S）的二聚化，在Vav的膜定位和激活中发挥重要作用。解决了VavS与GrbS配合物的晶体结构问题。VavS与其他SH 3结构域蛋白的不同之处在于其富含脯氨酸的肽的结合位点被其自身的RT环阻断。VavS β-桶的一端形成凹形疏水表面。GrbS组件与VavS表面形成连续的互补界面。GrbS对VavS的结合位点与富含脯氨酸的肽的典型结合位点部分重叠，但肯定不同。界面处的突变导致VavS对GrbS的结合亲和力降低4至40倍。该结构揭示了GrbS如何将VavS特异性地与其他信号分子区分开来，而不与富含脯氨酸的基序结合。

项目成果

Kawasaki, M, Inagaki, F.: "Random PCR-Based screening for soluble domains using green fluorescent protein"Biochem. Biophys. Res. Commun.. 280, 3. 842-844 (2001)

Kawasaki, M, Inagaki, F.：“使用绿色荧光蛋白进行基于随机 PCR 的可溶性结构域筛选”Biochem。

Ogura, K.: "Solution structure of N-terminal SH3 domain of Vav and the recognition site for Grb2 C-terminal SH3 domain"J.Biomol.NMR. 22. 37-46 (2002)

Ogura, K.：“Vav N 端 SH3 结构域的溶液结构和 Grb2 C 端 SH3 结构域的识别位点”J.Biomol.NMR。

Terasawa, H.: "Structure and ligand recognition of the PB1 domain : A novel protein Modulebinding to the PC motif"The EMBO J.. 20. 3947-3956 (2001)

Terasawa, H.：“PB1 结构域的结构和配体识别：与 PC 基序结合的新型蛋白质模块”EMBO J.. 20. 3947-3956 (2001)

Koga, H.: "Tetratricopeptide repeat (TPR) mitifs of p67 (phox) participate in interaction with the small GTPase Rc and activation of the phagocyte NADPH oxidase"J. Biol. Chem.. 274・35. 25051-25060 (1999)

Koga, H.：“p67 (phox) 的四肽重复序列 (TPR) 参与与小 GTP 酶 Rc 的相互作用和吞噬细胞 NADPH 氧化酶的激活”J. Biol. 274・35 (1999)。

Matsui,T.: "Rho-kinase phosphorylates COOH-terminal threonines of ezrin/radixin/moesin(ERM)proteins and regulates their head-to-tail association." J.Cell Biol.140. 647-657 (1998)

Matsui,T.：“Rho 激酶磷酸化 ezrin/radixin/moesin (ERM) 蛋白的 COOH 末端苏氨酸并调节它们的头尾关联。”