Fluorescence dynamics of green fluorescent protein in AOT reversed micelles.
We have used the enhanced green fluorescent protein (EGFP) to investigate the properties of surfactant-entrapped water pools in organic solvents (reversed micelles) with steady-state and time-resolved fluorescence methods. The surfactant used was sodium bis(2-ethylhexyl)sulfosuccinate (AOT) and the organic solvents were isooctane and (the more viscous) dodecane, respectively. The water content of the water pools could be controlled through the parameter w0, which is the water-to-surfactant molar ratio. With steady-state fluorescence, it was observed that subtle fluorescence changes could be noted in reversed micelles of different water contents. EGFP can be used as a pH-indicator of the water droplets in reversed micelles. Time-resolved fluorescence methods also revealed subtle changes in fluorescence decay times when the results in bulk water were compared with those in reversed micelles. The average fluorescence lifetimes of EGFP scaled with the relative fluorescence intensities. Time-resolved fluorescence anisotropy of EGFP in aqueous solution and reversed micelles yielded single rotational correlation times. Geometrical considerations could assign the observed correlation times to dehydrated protein at low w0 and internal EGFP rotation within the droplet at the highest w0.
Uskova
M A
MA
MicroSpectroscopy Centre, Department of Biomolecular Sciences, Wageningen University, The Netherlands.
Borst
J W
JW
Hink
M A
MA
van Hoek
A
A
Schots
A
A
Klyachko
N L
NL
Visser
A J
AJ
D000473:Alkanes / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D004143:Dioctyl Sulfosuccinic Acid / Q000378:metabolism* / Q000494:pharmacology; D005453:Fluorescence; D005454:Fluorescence Polarization; D049452:Green Fluorescent Proteins; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008164:Luminescent Proteins / Q000737:chemistry* / Q000378:metabolism; D008823:Micelles; D009795:Octanes / Q000378:metabolism / Q000494:pharmacology; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012399:Rotation; D012997:Solvents; D013050:Spectrometry, Fluorescence; D013501:Surface-Active Agents / Q000378:metabolism* / Q000494:pharmacology; D014867:Water / Q000378:metabolism / Q000494:pharmacology
D000473:Alkanes; D008164:Luminescent Proteins; D008823:Micelles; D009795:Octanes; D011994:Recombinant Proteins; D012997:Solvents; D013501:Surface-Active Agents; D014867:Water; D004143:Dioctyl Sulfosuccinic Acid; C007548:n-dodecane; D049452:Green Fluorescent Proteins; C045798:2,2,4-trimethylpentane