High stability of Discosoma DsRed as compared to Aequorea EGFP

Vladislav Verkhusha, Irina M. Kuznetsova, Olesia V. Stepanenko, Andrey G. Zaraisky, Michail M. Shavlovsky, Konstantin K. Turoverov, Vladimir N. Uversky

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish (US)
Pages (from-to)7879-7884
Number of pages6
JournalBiochemistry
Volume42
Issue number26
DOIs
StatePublished - Jul 8 2003
Externally publishedYes

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Embryonic Structures
Fluorescence
Cells
Proteins
Denaturation
Protein Stability
Guanidine
Chromophores
Xenopus
Insects
Cultured Cells
Rate constants
Hot Temperature
fluorescent protein 583
Degradation
Temperature
Population
enhanced green fluorescent protein
In Vitro Techniques

ASJC Scopus subject areas

  • Biochemistry

Cite this

Verkhusha, V., Kuznetsova, I. M., Stepanenko, O. V., Zaraisky, A. G., Shavlovsky, M. M., Turoverov, K. K., & Uversky, V. N. (2003). High stability of Discosoma DsRed as compared to Aequorea EGFP. Biochemistry, 42(26), 7879-7884. https://doi.org/10.1021/bi034555t

High stability of Discosoma DsRed as compared to Aequorea EGFP. / Verkhusha, Vladislav; Kuznetsova, Irina M.; Stepanenko, Olesia V.; Zaraisky, Andrey G.; Shavlovsky, Michail M.; Turoverov, Konstantin K.; Uversky, Vladimir N.

In: Biochemistry, Vol. 42, No. 26, 08.07.2003, p. 7879-7884.

Research output: Contribution to journalArticle

Verkhusha, V, Kuznetsova, IM, Stepanenko, OV, Zaraisky, AG, Shavlovsky, MM, Turoverov, KK & Uversky, VN 2003, 'High stability of Discosoma DsRed as compared to Aequorea EGFP', Biochemistry, vol. 42, no. 26, pp. 7879-7884. https://doi.org/10.1021/bi034555t
Verkhusha V, Kuznetsova IM, Stepanenko OV, Zaraisky AG, Shavlovsky MM, Turoverov KK et al. High stability of Discosoma DsRed as compared to Aequorea EGFP. Biochemistry. 2003 Jul 8;42(26):7879-7884. https://doi.org/10.1021/bi034555t
Verkhusha, Vladislav ; Kuznetsova, Irina M. ; Stepanenko, Olesia V. ; Zaraisky, Andrey G. ; Shavlovsky, Michail M. ; Turoverov, Konstantin K. ; Uversky, Vladimir N. / High stability of Discosoma DsRed as compared to Aequorea EGFP. In: Biochemistry. 2003 ; Vol. 42, No. 26. pp. 7879-7884.
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