Methods to examine the impact of nonsynonymous SNPs on protein degradation and function of human ABC transporter.
Methods to examine the impact of nonsynonymous SNPs on protein degradation and function of human ABC transporter.
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DOI：
10.1007/978-1-62703-435-7_15
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发表时间：
2013
影响因子：
--
通讯作者：
T. Ishikawa;Kanako Wakabayashi-Nakao;H. Nakagawa
中科院分区：
文献类型：
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作者：
T. Ishikawa;Kanako Wakabayashi-Nakao;H. Nakagawa
临床研究强烈表明，某些 ATP 结合盒 (ABC) 转运蛋白基因的遗传多态性和/或突变可能被视为影响患者对药物反应和/或疾病风险的重要因素。就 ABCG2 而言，编码基因中的某些单核苷酸多态性 (SNP) 会通过泛素介导的蛋白酶体蛋白水解途径改变底物特异性和/或增强从头合成的 ABCG2 蛋白的内质网相关降解 (ERAD)。迄今为止积累的临床数据表明，一些非同义 SNP 会影响 ABCG2 介导的药物或细胞代谢物的清除，尽管仍然存在一些争议。因此，我们最近开发了ABC转运蛋白的高速功能筛选和ERAD，以评估遗传多态性对其功能和体外蛋白表达水平的影响。在本章中，我们介绍了体外实验方法来阐明非同义 SNP 对 ABCG2 蛋白质降解及其转运功能的影响。
Clinical studies have strongly suggested that genetic polymorphisms and/or mutations of certain ATP-binding cassette (ABC) transporter genes might be regarded as significant factors affecting patients’ responses to medication and/or the risk of diseases. In the case of ABCG2, certain single nucleotide polymorphisms (SNPs) in the encoding gene alter the substrate specificity and/or enhance endoplasmic reticulum-associated degradation (ERAD) of the de novo synthesized ABCG2 protein via the ubiquitin-mediated proteasomal proteolysis pathway. Hitherto accumulated clinical data imply that several nonsynonymous SNPs affect the ABCG2-mediated clearance of drugs or cellular metabolites, although some controversies still exist. Therefore, we recently developed high-speed functional screening and ERAD of ABC transporters so as to evaluate the effect of genetic polymorphisms on their function and protein expression levels in vitro. In this chapter we present in vitro experimental methods to elucidate the impact of nonsynonymous SNPs on protein degradation of ABCG2 as well as on its transport function.