论文信息 - High stability of Discosoma DsRed as compared to Aequorea EGFP.

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.

[1] R. Tsien,et al. green fluorescent protein , 2020, Catalysis from A to Z.

[2] V. Subramaniam,et al. Oligomerization of DsRed is required for the generation of a functional red fluorescent chromophore , 2002, FEBS letters.

[3] Konstantin A Lukyanov,et al. A strategy for the generation of non‐aggregating mutants of Anthozoa fluorescent proteins , 2002, FEBS letters.

[4] Kei Ito,et al. An Enhanced Mutant of Red Fluorescent Protein DsRed for Double Labeling and Developmental Timer of Neural Fiber Bundle Formation* , 2001, The Journal of Biological Chemistry.

[5] D. Payan,et al. Characterization and Use of Green Fluorescent Proteins from Renilla mulleri and Ptilosarcus guernyi for the Human Cell Display of Functional Peptides , 2001, Journal of protein chemistry.

[6] M. Falk,et al. Expression of fluorescently tagged connexins: a novel approach to rescue function of oligomeric DsRed‐tagged proteins1 , 2001, FEBS letters.

[7] V. Verkhusha,et al. Denaturation and partial renaturation of a tightly tetramerized DsRed protein under mildly acidic conditions , 2000, FEBS letters.

[8] K. Mikoshiba,et al. A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications , 2002, Nature Biotechnology.

[9] A. Ciechanover,et al. The ubiquitin system for protein degradation. , 1992, Annual review of biochemistry.