Proteins on the move: insights gained from fluorescent protein technologies

• Published in: Nature reviews. Molecular cell biology Vol. 12; no. 10; pp. 656 - 668
• Main Author: Miyawaki, Atsushi
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.10.2011; Nature Publishing Group
• Subjects: 631/1647/1888/2249; 631/337; 81; 86; Animals; Biochemistry; Biology; Biomedical and Life Sciences; Biotechnology - methods; Cancer Research; Cell Biology; Cellular signal transduction; Developmental Biology; Genetic aspects; Green fluorescent protein; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Humans; Life Sciences; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; Physiological aspects; Protein Binding; Protein-protein interactions; Proteins; review-article; Signal transduction; Signal Transduction - genetics; Signal Transduction - physiology; Stem Cells; Japan
• ISSN: 1471-0072; 1471-0080
• DOI: 10.1038/nrm3199

Key Points In the past 10 years, there has been great progress in the development of fluorescent proteins (FPs) to study protein movement and protein interactions. Green FP (GFP)-like proteins have been mutated to be monomers as useful tags for analysing protein movement. FPs have been used to study bulk protein movement, primarily through photobleaching or photoactivation techniques, which allow the determination of protein diffusion and protein binding kinetics. FPs have also been used to track single proteins within the cell. The technique may vary depending on properties of the protein; for example, whether it is soluble. By studying movement, insights have been obtained on protein–protein interactions. Fluorescence cross-correlation spectroscopy (FCCS) has been used to detect the synchronous movement of two proteins fused to different FPs. Bimolecular fluorescence resonance energy transfer (FRET) and bimolecular fluorescence complementation (BiFC) have both been used to study protein proximity, an indicator of protein interactions. In the past 10 years, great progress has been made in the development of fluorescent proteins, including green fluorescent protein (GFP) and GFP-like proteins. Using these proteins together with a range of techniques has furthered our understanding of protein movement and protein–protein interactions. Proteins are always on the move, and this may occur through diffusion or active transport. The realization that the regulation of signal transduction is highly dynamic in space and time has stimulated intense interest in the movement of proteins. Over the past decade, numerous new technologies using fluorescent proteins have been developed, allowing us to observe the spatiotemporal dynamics of proteins in living cells. These technologies have greatly advanced our understanding of protein dynamics, including protein movement and protein interactions.