High stability of Discosoma DsRed as compared to Aequorea EGFP.
Comparative analysis of conformational stabilities was performed for two widely used genetic reporters, EGFP and DsRed, proteins exhibiting similar beta-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.
Verkhusha
Vladislav V
VV
Department of Pharmacology, University of Colorado Health Sciences Center, Denver, Colorado 80262, USA. Vlad.Verkhusha@uchsc.edu
Kuznetsova
Irina M
IM
Stepanenko
Olesia V
OV
Zaraisky
Andrey G
AG
Shavlovsky
Michail M
MM
Turoverov
Konstantin K
KK
Uversky
Vladimir N
VN
D000818:Animals; D002478:Cells, Cultured; D003063:Cnidaria / Q000737:chemistry*; D004331:Drosophila melanogaster / Q000378:metabolism; D005453:Fluorescence; D018507:Gene Expression Regulation, Developmental; D049452:Green Fluorescent Proteins; D019791:Guanidine / Q000494:pharmacology; D007202:Indicators and Reagents / Q000737:chemistry* / Q000378:metabolism; D008164:Luminescent Proteins / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D014982:Xenopus laevis / Q000196:embryology / Q000378:metabolism
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D011994:Recombinant Proteins; C413663:fluorescent protein 583; D049452:Green Fluorescent Proteins; D019791:Guanidine