Understanding FRET as a Research Tool for Cellular Studies

International Journal of Molecular Sciences

Dilip Shrestha¹  Attila Jenei¹  Péter Nagy¹  György Vereb¹  János Szöllősi¹ 

[1] Department of Biophysics and Cell Biology, University of Debrecen, Egyetem tér 1, Nagyerdei Krt. 98, Debrecen 4032, Hungary; E-Mails:

关键词: FRET;    Methods for measuring FRET;    Fluorescence intensity;    Fluorescence lifetime;    Anisotropy;    Major Histocompatibility Complex (MHC);    CD1d;    IL2;    IL15;    Immune synapse;    ErbB;   

DOI  :  10.3390/ijms16046718

来源: mdpi

【 摘 要 】

Communication of molecular species through dynamic association and/or dissociation at various cellular sites governs biological functions. Understanding these physiological processes require delineation of molecular events occurring at the level of individual complexes in a living cell. Among the few non-invasive approaches with nanometer resolution are methods based on Förster Resonance Energy Transfer (FRET). FRET is effective at a distance of 1–10 nm which is equivalent to the size of macromolecules, thus providing an unprecedented level of detail on molecular interactions. The emergence of fluorescent proteins and SNAP- and CLIP- tag proteins provided FRET with the capability to monitor changes in a molecular complex in real-time making it possible to establish the functional significance of the studied molecules in a native environment. Now, FRET is widely used in biological sciences, including the field of proteomics, signal transduction, diagnostics and drug development to address questions almost unimaginable with biochemical methods and conventional microscopies. However, the underlying physics of FRET often scares biologists. Therefore, in this review, our goal is to introduce FRET to non-physicists in a lucid manner. We will also discuss our contributions to various FRET methodologies based on microscopy and flow cytometry, while describing its application for determining the molecular heterogeneity of the plasma membrane in various cell types.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.