England Christopher G, Luo Haiming, Cai Weibo
†Department of Medical Physics, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.
‡Department of Radiology, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.
Bioconjug Chem. 2015 Jun 17;26(6):975-86. doi: 10.1021/acs.bioconjchem.5b00191. Epub 2015 May 22.
Exploration of protein function and interaction is critical for discovering links among genomics, proteomics, and disease state; yet, the immense complexity of proteomics found in biological systems currently limits our investigational capacity. Although affinity and autofluorescent tags are widely employed for protein analysis, these methods have been met with limited success because they lack specificity and require multiple fusion tags and genetic constructs. As an alternative approach, the innovative HaloTag protein fusion platform allows protein function and interaction to be comprehensively analyzed using a single genetic construct with multiple capabilities. This is accomplished using a simplified process, in which a variable HaloTag ligand binds rapidly to the HaloTag protein (usually linked to the protein of interest) with high affinity and specificity. In this review, we examine all current applications of the HaloTag technology platform for biomedical applications, such as the study of protein isolation and purification, protein function, protein-protein and protein-DNA interactions, biological assays, in vitro cellular imaging, and in vivo molecular imaging. In addition, novel uses of the HaloTag platform are briefly discussed along with potential future applications.
探索蛋白质功能和相互作用对于发现基因组学、蛋白质组学和疾病状态之间的联系至关重要;然而,生物系统中蛋白质组学的巨大复杂性目前限制了我们的研究能力。尽管亲和标签和自发荧光标签被广泛用于蛋白质分析,但这些方法的成功率有限,因为它们缺乏特异性,并且需要多个融合标签和基因构建体。作为一种替代方法,创新的卤代标签蛋白融合平台允许使用具有多种功能的单一基因构建体对蛋白质功能和相互作用进行全面分析。这通过一个简化的过程来实现,其中可变的卤代标签配体以高亲和力和特异性快速结合到卤代标签蛋白(通常与感兴趣的蛋白相连)上。在这篇综述中,我们研究了卤代标签技术平台目前在生物医学应用中的所有应用,如蛋白质分离和纯化研究、蛋白质功能、蛋白质-蛋白质和蛋白质-DNA相互作用、生物测定、体外细胞成像和体内分子成像。此外,还简要讨论了卤代标签平台的新用途以及潜在的未来应用。