title: In Vivo Analysis of Protein-Protein Interactions with Bioluminescence Resonance Energy Transfer (BRET): Progress and Prospects.
abstract: Proteins are the elementary machinery of life, and their functions are carried out mostly by molecular interactions. Among those interactions, protein-protein interactions (PPIs) are the most important as they participate in or mediate all essential biological processes. However, many common methods for PPI investigations are slightly unreliable and suffer from various limitations, especially in the studies of dynamic PPIs. To solve this problem, a method called Bioluminescence Resonance Energy Transfer (BRET) was developed about seventeen years ago. Since then, BRET has evolved into a whole class of methods that can be used to survey virtually any kinds of PPIs. Compared to many traditional methods, BRET is highly sensitive, reliable, easy to perform, and relatively inexpensive. However, most importantly, it can be done in vivo and allows the real-time monitoring of dynamic PPIs with the easily detectable light signal, which is extremely valuable for the PPI functional research. This review will take a comprehensive look at this powerful technique, including its principles, comparisons with other methods, experimental approaches, classifications, applications, early developments, recent progress, and prospects.
authors.affiliation: State Key Laboratory of Crop Stress Biology for Arid Areas and College of Life Sciences, Northwest A&F University, Yangling 712100, Shaanxi, China. sihuai.sun14@alumni.imperial.ac.uk.
authors.forename: Sihuai
authors.initials: S
authors.lastname: Sun
authors.affiliation: State Key Laboratory of Crop Stress Biology for Arid Areas and College of Life Sciences, Northwest A&F University, Yangling 712100, Shaanxi, China. bing3698@163.com.
authors.forename: Xiaobing
authors.initials: X
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Crop Stress Biology for Arid Areas and College of Life Sciences, Northwest A&F University, Yangling 712100, Shaanxi, China. wangyao@nwsuaf.edu.cn.
authors.forename: Yao
authors.initials: Y
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Crop Stress Biology for Arid Areas and College of Life Sciences, Northwest A&F University, Yangling 712100, Shaanxi, China. xihuishen@nwsuaf.edu.cn.
authors.forename: Xihui
authors.initials: X
authors.lastname: Shen
mesh_terms: D000818:Animals; D006801:Humans; D008156:Luciferases / Q000737:chemistry; D008163:Luminescent Measurements / Q000639:trends*; D011506:Proteins / Q000378:metabolism*
chemical_list: D011506:Proteins; D008156:Luciferases
keywords: Bioluminescence Resonance Energy Transfer; Förster Resonance Energy Transfer; NanoLuc; bacterial luciferase; bioluminescence; fluorescence; protein dynamics; protein–protein interactions