Department of Chemistry, Pennsylvania State University, University Park, Pennsylvania, PA 16802, USA.
Chembiochem. 2020 Jul 16;21(14):1935-1946. doi: 10.1002/cbic.202000037. Epub 2020 Apr 2.
Visualizing and manipulating the behavior of proteins is crucial to understanding the physiology of the cell. Methods of biorthogonal protein labeling are important tools to attain this goal. In this review, we discuss advances in probe technology specific for self-labeling protein tags, focusing mainly on the application of HaloTag and SNAP-tag systems. We describe the latest developments in small-molecule probes that enable fluorogenic (no wash) imaging and super-resolution fluorescence microscopy. In addition, we cover several methodologies that enable the perturbation or manipulation of protein behavior and function towards the control of biological pathways. Thus, current technical advances in the HaloTag and SNAP-tag systems means that they are becoming powerful tools to enable the visualization and manipulation of biological processes, providing invaluable scientific insights that are difficult to obtain by traditional methodologies. As the multiplex of self-labeling protein tag systems continues to be developed and expanded, the utility of these protein tags will allow researchers to address previously inaccessible questions at the forefront of biology.
可视化和操纵蛋白质的行为对于理解细胞的生理学至关重要。生物正交蛋白质标记方法是实现这一目标的重要工具。在这篇综述中,我们讨论了专门用于自标记蛋白质标签的探针技术的进展,主要集中在 HaloTag 和 SNAP-tag 系统的应用上。我们描述了可用于荧光(无需洗涤)成像和超分辨率荧光显微镜的小分子探针的最新进展。此外,我们还介绍了几种能够干扰或操纵蛋白质行为和功能以控制生物途径的方法。因此,HaloTag 和 SNAP-tag 系统的当前技术进步意味着它们正在成为可视化和操纵生物过程的强大工具,为生物学前沿提供了难以通过传统方法获得的宝贵科学见解。随着自标记蛋白质标签系统的多路复用不断发展和扩展,这些蛋白质标签的实用性将使研究人员能够解决生物学前沿领域以前无法解决的问题。