期刊
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
卷 16, 期 4, 页码 6718-6756出版社
MDPI
DOI: 10.3390/ijms16046718
关键词
FRET; Methods for measuring FRET; Fluorescence intensity; Fluorescence lifetime; Anisotropy; Major Histocompatibility Complex (MHC); CD1d; IL2; IL15; Immune synapse; ErbB
资金
- Hungarian Scientific Research Fund [NK 101337, K103906]
- European Commission [(LSHC-CT-2005-018914)-ATTACK, MCRTN-CT-2006-036946-2]
- New Hungary Development Plan - European Social Fund
- European Regional Development Fund [TAMOP-4.2.2.A-11/1/KONV-2012-0025, TAMOP-4.2.2-08/1-2008-0019, TAMOP-4.2.1/B-09/1/KONV-2010-007]
- Baross Gabor Program [REG-EA-09-1-2009-0010]
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 Forster 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.
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