蛋白激酶与磷酸酶.PPT

EEVAS*EPEEAAS*PTTPK EEVAS*EPEEAASPTTPK EEVASEPEEAASPTTPK y1-y5, M y6-y12, M+80 y13-y17,M+160 y1-y17, M y1-y12, M y13-y17,M+80 生物质谱分析蛋白质的磷酸化修饰 IMAC: Immobilized Metal Affinity Chromatography 固定化金属离子亲和性层析法 SIMAC: sequential elution from IMAC 从IMAC顺序洗脱 磷酸化蛋白质组（Phosphoproteome） DUSP1 PI3K信号通路与癌症治疗 PI3K信号通路参与调控细胞的生长、存活、代谢等重要的生命活动； PI3K激酶在多数癌变组织中激活，而其催化亚基p110α在大约15%的癌细胞中发生激活突变或基因扩增； 其特异性的磷酸酶PTEN是癌细胞中最常发生突变的抑癌基因之一，仅次于p53； PI3K激酶上下游分子如受体酪氨酸激酶、Akt激酶等的激活也可以促进癌症的发生与发展。 PI3K信号通路与癌症治疗 * GF: such as EGF, TGF, PDGF etc. Mutated ras Schematic representation of c-Src activation at the onset of mitosis. (A) During most of the cell cycle, c-Src is largely phosphorylated at Tyr527 and therefore in an inactive tail-bite conformation, whereas 20% of RPTP is phosphorylated at Tyr789 and saturated with Grb2. (B) At the transition from G2 to M phase, c-Src is first phosphorylated in its NH2-terminus by p34cdc2, and RPTP is phosphorylated at several serines by an unknown kinase, perhaps PKC. Subsequently, Grb2 dissociates from RPTP . (C) c-Src binds through its SH2 domain to phospho-Tyr789 on RPTP . This binding event exposes phospho-Tyr527 for dephosphorylation by the D1 domain of RPTP , allowing c-Src to autophosphorylate at Tyr416, thereby becoming fully active. PTK, protein tyrosine kinase. * 1. Wortmannin is a reactive electrophilic and therefore unstable compound whereas LY294002 is a weak inhibitor with only micromolar potency, and both agents display significant off-target effects on other kinases。 蛋白激酶与磷酸酶 病理生理学教研室 蛋白质翻译后修饰 Post-translational modifications (PTMs) 磷酸化修饰（Phosphorylation ） 糖基化修饰（Glycosylation ） 泛素化修饰（Ubiquitination ） 乙酰化修饰（Acetylation ） 甲基化修饰（Methylation ） SUMOylation…… 磷酸化修饰的氨基酸 蛋白质的磷酸化修饰 导致受体蛋白的激活或失活； 2% 蛋白质的磷酸化修饰发生在Tyr残基上； 大部分的早期信号转导是由酪氨酸蛋白激酶介导的（protein tyrosine kinase, PTKs)； Ser/Thr磷酸化修饰多发生于信号转导级联反应的下游。 蛋白激酶将ATP末端的磷酸基团转移到蛋白质侧链的羟基上； 磷酸酶催化蛋白质侧链上磷酸基团的水解。 蛋白激酶与磷酸酶 ~30%的蛋白质可以发生磷酸化修饰 人类的激酶组包括518蛋白激酶基因 其中218个基因与人类疾病的发生发展密切相关 约30个基因是肿瘤抑制基因 约100个为原癌基因 Hu