BCR-ABL与泛素E3连接酶c-CBL的相互作用在慢性髓系白血病靶向治疗中的作用及相关机制研究

发布时间：2018-04-05 13:08

本文选题：BCR-ABL　切入点：c-CBL　出处：《中国实验血液学杂志》2017年01期

【摘要】：目的:通过对BCR-ABL与泛素E3连接酶c-CBL的相互作用结构域的筛查,明确砷剂治疗慢性髓系白血病(CML)的结构基础。方法:根据BCR-ABL与c-CBL的结构信息,对二者的结合界面进行结构模拟与分析,基于此构建BCR-ABL的不同突变体[(△SH2(Src同源结构域2)、△TyrKC(酪氨酸激酶催化结构域)和△SH2/TyrKC(S/H))以及c-CBL的突变体△RF(环指结构域),转染293T及HeLa细胞,应用免疫共沉淀(Co-IP)]及免疫荧光(IF)共定位的方法筛查BCR-ABL与c-CBL的相互作用结构域。结果:Co-IP发现BCR-ABL的TyrKC结构域主要参与了与c-CBL的相互作用,SH2结构域也发挥一定的作用,但作用弱于TyrKC结构域;当两个结构域同时截短后,BCR-ABL与c-CBL几乎不发生相互作用;同时c-CBL的RF结构域在一定程度上影响了与BCR-ABL的相互作用。IF的结果与Co-IP相符,证明了结构模拟的准确性。结论:BCR-ABL的TyrKC和SH2结构域主要参与了与c-CBL的相互作用,而c-CBL的RF结构域参与了与BCR-ABL的相互作用。c-CBL与BCR-ABL的相互作用对BCR-ABL的稳定性发挥调控作用,对于深入理解砷剂靶向治疗CML分子机制提供了结构基础。
[Abstract]:Aim: to screen the interaction domain of BCR-ABL with ubiquitin E3 ligase c-CBL and to clarify the structural basis of arsenic in the treatment of chronic myeloid leukemia (CML).Methods: according to the structure information of BCR-ABL and c-CBL, the structure of the interface was simulated and analyzed.Different mutants of BCR-ABL [(SH2(Src homologous domain 2N, TyrKC (tyrosine kinase catalytic domain)) and SH2 / TyrKC- S- / HN] and c-CBL mutant RF- (ring finger domain) were constructed based on these mutants, which were transfected into 293T and HeLa cells.The interaction domain between BCR-ABL and c-CBL was screened by co-localization of co-precipitation and immunofluorescence.Results the TyrKC domain of BCR-ABL was found to play a role in the interaction with c-CBL, but the effect was weaker than that of TyrKC domain, and when the two domains were truncated at the same time, there was almost no interaction between BCR-ABL and c-CBL.At the same time, the RF domain of c-CBL affects the interaction with BCR-ABL to some extent. The results of if agree with that of Co-IP, which proves the accuracy of structural simulation.Conclusion the TyrKC and SH2 domains of BCR-ABL participate in the interaction with c-CBL, while the RF domain of c-CBL participates in the interaction with BCR-ABL. The interaction between c-CBL and BCR-ABL plays a regulatory role in the stability of BCR-ABL.It provides a structural basis for further understanding the molecular mechanism of arsenic targeted therapy for CML.
【作者单位】：医学基因组学国家重点实验室上海血液学研究所上海交通大学医学院附属瑞金医院;药物研究国家重点实验室药物研发中心中国科学院上海药物研究所;
【基金】：国家重大基础研究计划项目973项目(2013CB966800) 国家自然科学基金(81670127,81270594,81101721) 上海市浦江人才计划项目(16PJ1406100) 上海市自然科学基金项目(16ZR1421000) 上海市青年科技启明星项目(13QA1402600) 教育部高等学校博士学科点专项科研基金项目(新教师类)(20100073120095)
【分类号】：R733.72

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