Fluorescence resonance energy transfer (FRET) applications using green fluorescent protein. Energy transfer to the endogenous chromophores of phycobilisome light-harvesting complexes
Van Thor, J.J.; Hellingwerf, K.J.
Methods in Molecular Biology 183: 101-119
2002
ISSN/ISBN: 1064-3745 PMID: 12136747 DOI: 10.1385/1-59259-280-5:101
Accession: 046106124
Fluorescence resonance energy transfer (FRET) is a technique that can be used to estimate intermolecular distances between pigment molecules, which is an approach first proposed by Stryer (1). The theoretical basis for Fret was originally put forward by Förster (2,3), and is related to "Fermi's golden rule" for electronic interactions. This chapter explains how and when Fret can be used, and what the physical basis is of the energy transfer events. The point is stressed that, in most cases, Fret cannot be used to directly measure intermolecular distances, but only to estimate them. The reason for these uncertainties are made clear. First, the mechanisms of fluorescence and fluorescence energy transfer are briefly introduced.