Fluorescence lifetime imaging of coral fluorescent proteins.
Corals, like many other coelenterates, contain fluorescent pigments that show considerable homology with the well known green fluorescent protein of the jellyfish Aequoria. In corals, unlike jellyfish, multiple proteins are present and the range of excitations and emissions suggest the possibility of energy transfer. The occurrence of Förster resonant energy transfer (FRET) between fluorescent proteins in corals has already been reported and time-resolved spectra have shown the effect on fluorescent lifetime, but without any spatial resolution. Lifetime confocal microscopy offers lower time resolution but excellent spatial resolution. Lifetimes of the isolated A. millepora pigments amilFP490, amilFP504, and amilFP593 (names indicate emission peaks) were 2.8, 2.9, and 2.9 ns, respectively. In the coral sample, imaging the entire emission spectrum from 420 nm, the mean lifetime was reduced to 1.5 ns, implying that FRET was occurring. Looking just at the fluorescence from FRET donors the lifetime was even shorter, at 1.3 ns, supporting this interpretation. In contrast, no reduction in lifetime is seen in the coral Euphyllia ancora, where the pigment distribution also suggests that the pigments are unlikely to be involved in photoprotection. This study set out to determine the extent of FRET between pigments in two corals, Acropora millepora and Euphyllia, ancora which differ in the arrangement of their pigments and hence possibly in pigment function.
Cox
Guy
G
Australian Key Centre for Microscopy and Microanalysis, The University of Sydney, Sydney, New South Wales, Australia. guy.cox@emu.usyd.edu.au
Matz
Mikhail
M
Salih
Anya
A
D000818:Animals; D037421:Anthozoa / Q000378:metabolism*; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D049452:Green Fluorescent Proteins / Q000032:analysis; D008164:Luminescent Proteins / Q000032:analysis*; D008856:Microscopy, Fluorescence / Q000379:methods*; D010860:Pigments, Biological / Q000032:analysis
D008164:Luminescent Proteins; D010860:Pigments, Biological; D049452:Green Fluorescent Proteins