机构地区:[1]中国科学院生态环境研究中心,环境化学与生态毒理学国家重点实验室,北京100085 [2]中国科学院大学,北京100049
出 处:《色谱》2020年第10期1133-1142,共10页Chinese Journal of Chromatography
基 金:国家自然科学基金项目(91743201,21621064)。
摘 要:蛋白质-DNA的相互作用在决定细胞命运的许多过程中发挥重要作用,对蛋白质-DNA相互作用的分子机制研究有利于对基本生命过程的理解,为相关疾病的临床治疗及药物筛选提供理论指导。另一方面,利用一些已知的蛋白质-DNA相互作用可以帮助开发先进的生物工程和生命分析技术,为相关研究提供有力的技术支持。因此,建立灵敏、快速的分析方法用于表征蛋白质-DNA的相互作用十分重要。高效毛细管电泳(capillary electrophoresis,CE)技术因其超高的分离效率、极低的样品消耗与较短的分析时间等优势被广泛应用于化学、生命科学和环境科学等多个研究领域。其中,亲和毛细管电泳(affinity capillary electrophoresis,ACE)技术已经成为考察分子间相互作用的重要研究工具。这篇文章综述了亲和毛细管电泳技术自建立以来在蛋白质-DNA相互作用分析方面的研究进展,并对经典的研究工作进行了着重介绍,主要包括三方面的内容:(1)亲和毛细管电泳技术简介;(2)利用亲和毛细管电泳技术进行蛋白质-DNA相互作用的基础分子机制研究;(3)利用已知的蛋白质-DNA相互作用发展针对目标分子及目标反应的亲和毛细管电泳检测技术。本文还对该领域的未来发展趋势进行了展望与探讨,提出应从以下两个方面增强亲和毛细管电泳技术的分析能力:(1)充分发挥CE技术样品消耗少和高通量等优势,分别发展针对少量珍贵生物样品的高灵敏检测方法和针对大量未知因素的高通量筛选方法;(2)结合DNA测序及质谱技术快速筛选、鉴定未知的蛋白质-DNA相互作用的精确靶点。Protein-DNA interactions play essential roles in various biological events that determine the cell fate.Research on the molecular mechanism of protein-DNA interactions has helped elucidate diverse fundamental life processes,thereby providing theoretical guidance for establishing clinical treatment and screening potential drug of target diseases.Furthermore,well-known protein-DNA interactions have been utilized to develop advanced bioengineering and bioanalytical techniques,therefore providing robust technical support for related research.Hence,it is important to establish sensitive and rapid analytical methods to study protein-DNA interactions.High-performance capillary electrophoresis(CE)has been widely used in many research fields such as chemistry,life sciences,and environmental sciences,mainly due to its advantages including ultra-high separation efficiency,extremely low sample consumption,and short analysis time.For instance,affinity capillary electrophoresis(ACE)has become an important analytical tool for investigating molecular interactions.In this paper,we review the applications of ACE in studying protein-DNA interactions since it was first proposed in 1992,addressing previous significant work in this field.Three major aspects have been summarized in this review:(1)brief introduction to the development of ACE technique;(2)applications of ACE in the fundamental research on the molecular mechanism of protein-DNA interactions;and(3)applications of well-known protein-DNA interactions in CE-based detection of target molecules and reactions.In the first aspect,along with the concept and separation modes of ACE,general strategies to enhance the analytical ability of ACE are briefly introduced.In the second aspect,the applications of ACE in studying several important protein-DNA interactions involving transcription factors(e.g.,GCN4),DNA repair proteins(e.g.,UvrA,UvrB,and RecA),and methylated DNA-binding proteins(MBDs)are reviewed.In the third aspect,the applications of well-known molecular interactions(e.g.,ant
关 键 词:亲和毛细管电泳 蛋白质-DNA相互作用 亲和分析 综述
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