Effect of high pressure and reversed micelles on the fluorescent proteins.
Two physico-chemical perturbations were applied to ECFP, EGFP, EYFP and DsRed fluorescent proteins: high hydrostatic pressure and encapsulation in reversed micelles. The observed fluorescence changes were described by two-state model and quantified by thermodynamic formalism. ECFP, EYFP and DsRed exhibited similar reaction volumes under pressure. The changes of the chemical potentials of the chromophore in bis(2-ethylhexyl)sulfosuccinate (AOT) micelles caused apparent chromophore protonation changes resulting in a fluorescence decrease of ECFP and EYFP. In contrast to the remarkable stability of DsRed, the highest sensitivity of EYFP fluorescence under pressure and in micelles is attributed to its chromophore structure.
Verkhusha
Vladislav V
VV
Department of Pharmacology, C236, University of Colorado Health Sciences Center, 4200 East Ninth Avenue, Denver, CO 80262, USA. vlad.verkhusha@uchsc.edu
Pozhitkov
Alexander E
AE
Smirnov
Sergey A
SA
Borst
Jan Willem
JW
van Hoek
Arie
A
Klyachko
Natalya L
NL
Levashov
Andrey V
AV
Visser
Antonie J W G
AJ
D004355:Drug Stability; D005453:Fluorescence; D049452:Green Fluorescent Proteins; D006874:Hydrostatic Pressure; D008164:Luminescent Proteins / Q000737:chemistry*; D008823:Micelles; D013816:Thermodynamics
D008164:Luminescent Proteins; D008823:Micelles; C413663:fluorescent protein 583; D049452:Green Fluorescent Proteins