Novel Insights in Membrane Biology Utilizing Fluorescence Recovery After Photobleaching
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Understanding cellular signaling by membrane receptors in terms of their lateral dynamics represents a challenging area in contemporary biology. Fluorescence recovery after photobleaching (FRAP) offers a convenient approach to measure lateral diffusion and is extensively used for measuring lateral diffusion of lipids and proteins in membranes. In this review, we have provided an overview of the type of questions that could be addressed in membrane and receptor biology utilizing FRAP, with representative examples chosen from work carried out in our group. A major focus is on exploring new horizons in the organization and dynamics of G protein-coupled receptors (GPCRs) utilizing FRAP. We discuss how lateral dynamics of membrane receptors could serve as crucial determinants of their signaling. We envision that FRAP, along with confocal microscopy, could provide novel insight into dynamics of intracellular organelles.
- 5-HT1A receptor
5-Hydroxytryptamine-1A receptor tagged to enhanced yellow fluorescent protein
Enhanced yellow fluorescent protein
- FAST DiI
Fluorescence recovery after photobleaching
Green fluorescent protein
G protein-coupled receptor
We dedicate this paper to Prof. Michael Edidin (The Johns Hopkins University, Baltimore, MD) who pioneered the application of FRAP in biological membranes and in whose laboratory one of us (A.C.) learnt the nuts and bolts of FRAP measurements during a visit as a CSIR-Raman Fellow. Work in A.C.’s laboratory was supported by the Council of Scientific and Industrial Research, Govt. of India. A.C. is an Adjunct Professor at the Special Centre for Molecular Medicine of Jawaharlal Nehru University (New Delhi, India) and Indian Institute of Science Education and Research (Mohali, India), and Honorary Professor of the Jawaharlal Nehru Centre for Advanced Scientific Research (Bangalore, India). A.C. gratefully acknowledges J.C. Bose Fellowship (Dept. of Science and Technology, Govt. of India). Some of the work described in this article was carried out by former members of A.C.’s group whose contributions are gratefully acknowledged. We thank members of our laboratory for critically reading the manuscript.
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