本文選題：BCR-ABL　切入點：c-CBL　出處：《中國實驗血液學(xué)雜志》2017年01期

【摘要】：目的:通過對BCR-ABL與泛素E3連接酶c-CBL的相互作用結(jié)構(gòu)域的篩查,明確砷劑治療慢性髓系白血病(CML)的結(jié)構(gòu)基礎(chǔ)。方法:根據(jù)BCR-ABL與c-CBL的結(jié)構(gòu)信息,對二者的結(jié)合界面進(jìn)行結(jié)構(gòu)模擬與分析,基于此構(gòu)建BCR-ABL的不同突變體[(△SH2(Src同源結(jié)構(gòu)域2)、△TyrKC(酪氨酸激酶催化結(jié)構(gòu)域)和△SH2/TyrKC(S/H))以及c-CBL的突變體△RF(環(huán)指結(jié)構(gòu)域),轉(zhuǎn)染293T及HeLa細(xì)胞,應(yīng)用免疫共沉淀(Co-IP)]及免疫熒光(IF)共定位的方法篩查BCR-ABL與c-CBL的相互作用結(jié)構(gòu)域。結(jié)果:Co-IP發(fā)現(xiàn)BCR-ABL的TyrKC結(jié)構(gòu)域主要參與了與c-CBL的相互作用,SH2結(jié)構(gòu)域也發(fā)揮一定的作用,但作用弱于TyrKC結(jié)構(gòu)域;當(dāng)兩個結(jié)構(gòu)域同時截短后,BCR-ABL與c-CBL幾乎不發(fā)生相互作用;同時c-CBL的RF結(jié)構(gòu)域在一定程度上影響了與BCR-ABL的相互作用。IF的結(jié)果與Co-IP相符,證明了結(jié)構(gòu)模擬的準(zhǔn)確性。結(jié)論:BCR-ABL的TyrKC和SH2結(jié)構(gòu)域主要參與了與c-CBL的相互作用,而c-CBL的RF結(jié)構(gòu)域參與了與BCR-ABL的相互作用。c-CBL與BCR-ABL的相互作用對BCR-ABL的穩(wěn)定性發(fā)揮調(diào)控作用,對于深入理解砷劑靶向治療CML分子機(jī)制提供了結(jié)構(gòu)基礎(chǔ)。
[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.
【作者單位】： 醫(yī)學(xué)基因組學(xué)國家重點實驗室上海血液學(xué)研究所 上海交通大學(xué)醫(yī)學(xué)院附屬瑞金醫(yī)院;藥物研究國家重點實驗室藥物研發(fā)中心中國科學(xué)院上海藥物研究所;
【基金】：國家重大基礎(chǔ)研究計劃項目973項目(2013CB966800) 國家自然科學(xué)基金(81670127,81270594,81101721) 上海市浦江人才計劃項目(16PJ1406100) 上海市自然科學(xué)基金項目(16ZR1421000) 上海市青年科技啟明星項目(13QA1402600) 教育部高等學(xué)校博士學(xué)科點專項科研基金項目(新教師類)(20100073120095)
【分類號】：R733.72

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