TY - JOUR
T1 - High stability of Discosoma DsRed as compared to Aequorea EGFP
AU - Verkhusha, Vladislav V.
AU - Kuznetsova, Irina M.
AU - Stepanenko, Olesia V.
AU - Zaraisky, Andrey G.
AU - Shavlovsky, Michail M.
AU - Turoverov, Konstantin K.
AU - Uversky, Vladimir N.
N1 - Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2003/7/8
Y1 - 2003/7/8
N2 - Comparative analysis of conformational stabilities was performed for two widely used genetic reporters, EGFP and DsRed, proteins exhibiting similar β-can folds, but possessing different oligomeric organization and chromophore structures. Two factors affecting protein stability in vitro, such as elevated temperatures and a chaotropic agent guanidine hydrochloride, were studied. In vivo tolerance of the fluorescence proteins to proteasomal-based degradation was studied in insect and mammalian cells, and in Xenopus embryos. The apparent rate constants of thermal and GdmCl-induced denaturation were several orders of magnitude lower for DsRed than for EGFP. DsRed lifetimes severalfold longer than those of EGFP were observed in cultured cells and in embryos. The remarkable fluorescence stability of DsRed under the all conditions that have been studied is attributed to a significant extent to its tetrameric organization. Therefore, DsRed can be used as a genetic reporter and advanced population marker with a significantly extended intracellular lifespan.
AB - Comparative analysis of conformational stabilities was performed for two widely used genetic reporters, EGFP and DsRed, proteins exhibiting similar β-can folds, but possessing different oligomeric organization and chromophore structures. Two factors affecting protein stability in vitro, such as elevated temperatures and a chaotropic agent guanidine hydrochloride, were studied. In vivo tolerance of the fluorescence proteins to proteasomal-based degradation was studied in insect and mammalian cells, and in Xenopus embryos. The apparent rate constants of thermal and GdmCl-induced denaturation were several orders of magnitude lower for DsRed than for EGFP. DsRed lifetimes severalfold longer than those of EGFP were observed in cultured cells and in embryos. The remarkable fluorescence stability of DsRed under the all conditions that have been studied is attributed to a significant extent to its tetrameric organization. Therefore, DsRed can be used as a genetic reporter and advanced population marker with a significantly extended intracellular lifespan.
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U2 - 10.1021/bi034555t
DO - 10.1021/bi034555t
M3 - Article
C2 - 12834339
AN - SCOPUS:0037710130
SN - 0006-2960
VL - 42
SP - 7879
EP - 7884
JO - Biochemistry
JF - Biochemistry
IS - 26
ER -