项目名称: 利用金属蛋白质组学研究砷结合蛋白及其在砷皮肤毒性中的作用
项目编号: No.21507153
项目类型: 青年科学基金项目
立项/批准年度: 2016
项目学科: 化学工业
项目作者: 郭志灵
作者单位: 中国科学院生态环境研究中心
项目金额: 21万元
中文摘要: 我国是砷污染最严重的国家之一,地方性砷中毒引起的皮肤癌患者越来越多。由于砷毒性机制仍不清楚,目前对砷中毒尚无有效的预防和治疗措施。砷与蛋白质的相互作用是了解砷毒性机制的关键所在。目前,砷在皮肤中与哪些蛋白结合以及这些砷结合蛋白在砷毒性中的作用尚不明确。常规的砷化学形态分析方法需要复杂的样品处理,且无法原位分析。本项目将以人永生化角质形成细胞为研究对象,以亚砷酸钠为砷污染物的代表,利用基于同步辐射的金属蛋白质组学技术对砷在亚细胞水平的分布和化学种态进行分析,通过同步辐射微束X射线荧光与二维凝胶电泳准在线联用,找出砷结合蛋白,并利用基因沉默与过表达等技术探讨关键砷结合蛋白在砷毒性中的作用。该项目的成果将为进一步理解砷的皮肤毒性机制以及砷中毒的早期诊断和治疗提供新的重要的科学依据。
中文关键词: 砷污染;结合蛋白;金属蛋白质组学;同步辐射;角化细胞
英文摘要: China is among the countries which have been severely contaminated by arsenic (As). Arsenic-associated skin lesions and cancers are readily increasing in arsenic-exposed individuals. However, the molecular mechanisms of arsenic-induced cancers are still unknown, and there are still no effective treatment options. Binding of arsenic with protein is considered as a key factor for understanding the molecular mechanisms of arsenic toxicity. Till now, As-binding proteins in skin and its role in As-induced toxicity are not clear. Traditional speciation analysis for arsenic in biological samples requires complex sample processing, and cannot obtain the in situ information. This project will use human spontaneously immortalized skin keratinocytes (HaCaT) as cell model and NaAsO2 as representative arsenic contaminant, aims to: 1) investigate the subcellular distribution and speciation of arsenic in situ, 2) screen out arsenic-binding proteins by synchrotron-based metalloproteomics, 3) and explore the role of arsenic-binding proteins in arsenic induced toxicity by gene knock-down and overexpression techniques. This project will provide important scientific basis for understanding the mechanism of arsenic-induced toxicity and early diagnosis and treatment for arseniasis.
英文关键词: arsenic pollution;binding protein;metalloproteomics;synchrotron radiation;keratinocyte