Lunev Sergey, Semmelink Marije F W, Xian Jia Ling, Ma Kai Yu, Leenders Anna J A, Dömling Alexander S S, Shtutman Michael, Groves Matthew R
Department of Drug Design, Groningen Research Institute of Pharmacy, University of Groningen, Antonius Deusinglaan 1, 9700 AD Groningen, The Netherlands.
Department of Drug Discovery and Biomedical Sciences, South Carolina College of Pharmacy, University of South Carolina, 715 Sumter Street, Columbia, SC 29208, USA.
Acta Crystallogr F Struct Biol Commun. 2017 Jan 1;73(Pt 1):1-8. doi: 10.1107/S2053230X16018896.
The majority of modern anticancer approaches target DNA/protein targets involved in tumour-cell proliferation. Such approaches have a major drawback, as nonproliferating cancer cells remain unaffected and may cause relapse or remission. Human coatomer protein complex I (COPI) subunit ζ (Copζ), a component of the coat protein involved in cell apoptosis and intracellular trafficking, has recently been proposed as a potential anticancer drug target. Previous studies have shown that two different isoforms of the Copζ subunit exist in mammalian cells. While normal cells express both Copζ1 and Copζ2 isoforms, various types of tumour cells display a loss of Copζ2 expression and rely solely on Copζ1 for growth and survival. Subsequent knockdown of Copζ1 results in specific inhibition of both proliferating and dormant tumour-cell populations, with no adverse growth effects on normal cells. Therefore, a Copζ1-targeting therapy was proposed to bypass the problem of dormant cancer cells that are resistant to conventional antiproliferative drugs, which is the major cause of tumour relapse. In order to aid in structure-based inhibitor design, a crystal structure is required. In this article, the recombinant expression, purification, crystallization and crystal structure of Copζ1, as well as the expression and purification of Copζ2, are reported.
大多数现代抗癌方法都针对参与肿瘤细胞增殖的DNA/蛋白质靶点。这类方法有一个主要缺点,即非增殖性癌细胞不受影响,可能导致复发或缓解。人衣被蛋白复合物I(COPI)亚基ζ(Copζ)是参与细胞凋亡和细胞内运输的衣被蛋白的一个组成部分,最近被提议作为一种潜在的抗癌药物靶点。先前的研究表明,哺乳动物细胞中存在两种不同的Copζ亚基同工型。正常细胞同时表达Copζ1和Copζ2同工型,而各种类型的肿瘤细胞则表现出Copζ2表达缺失,仅依赖Copζ1进行生长和存活。随后敲低Copζ1会导致增殖性和休眠性肿瘤细胞群体受到特异性抑制,而对正常细胞没有不良生长影响。因此,有人提出一种靶向Copζ1的疗法,以绕过对传统抗增殖药物有抗性的休眠癌细胞问题,而这正是肿瘤复发的主要原因。为了辅助基于结构的抑制剂设计,需要晶体结构。本文报道了Copζ1的重组表达、纯化、结晶和晶体结构,以及Copζ2的表达和纯化。