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Single amino acid replacement makes Aequorea victoria fluorescent proteins reversibly photoswitchable.
J Am Chem Soc 2010; 132(1):85-95JA

Abstract

Reversibly photoswitchable (i.e., photochromic) fluorescent proteins open the way to a number of advanced bioimaging techniques applicable to living-cell studies such as sequential photolabeling of distinct cellular regions, innovative FRET schemes, or nanoscopy. Owing to the relevance of fluorescent proteins from Aequorea victoria (AFPs) for cell biology, a photochromic "toolbox" constituted by several AFPs is highly desirable. Here we introduce four new photochromic AFPs whose reversible photoswitching occurs between the native bright and a dark state at low illumination power, on account of a very efficient cis-trans photoisomerization. Most remarkably, the optical bistability of these AFPs derives from the single E222Q mutation in the primary sequence. Apparently, the E222Q substitution can restore the intrinsic photochromic behavior of the isolated chromophore. The significance of these mutants for high-resolution in vivo cell imaging is shown by means of photochromic FRET experiments.

Authors+Show Affiliations

IIT@NEST, Center for Nanotechnology Innovation, Piazza San Silvestro 12, I-56127 Pisa, Italy. r.bizzarri@sns.itNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19958004

Citation

Bizzarri, Ranieri, et al. "Single Amino Acid Replacement Makes Aequorea Victoria Fluorescent Proteins Reversibly Photoswitchable." Journal of the American Chemical Society, vol. 132, no. 1, 2010, pp. 85-95.
Bizzarri R, Serresi M, Cardarelli F, et al. Single amino acid replacement makes Aequorea victoria fluorescent proteins reversibly photoswitchable. J Am Chem Soc. 2010;132(1):85-95.
Bizzarri, R., Serresi, M., Cardarelli, F., Abbruzzetti, S., Campanini, B., Viappiani, C., & Beltram, F. (2010). Single amino acid replacement makes Aequorea victoria fluorescent proteins reversibly photoswitchable. Journal of the American Chemical Society, 132(1), pp. 85-95. doi:10.1021/ja9014953.
Bizzarri R, et al. Single Amino Acid Replacement Makes Aequorea Victoria Fluorescent Proteins Reversibly Photoswitchable. J Am Chem Soc. 2010 Jan 13;132(1):85-95. PubMed PMID: 19958004.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Single amino acid replacement makes Aequorea victoria fluorescent proteins reversibly photoswitchable. AU - Bizzarri,Ranieri, AU - Serresi,Michela, AU - Cardarelli,Francesco, AU - Abbruzzetti,Stefania, AU - Campanini,Barbara, AU - Viappiani,Cristiano, AU - Beltram,Fabio, PY - 2009/12/5/entrez PY - 2009/12/5/pubmed PY - 2010/3/2/medline SP - 85 EP - 95 JF - Journal of the American Chemical Society JO - J. Am. Chem. Soc. VL - 132 IS - 1 N2 - Reversibly photoswitchable (i.e., photochromic) fluorescent proteins open the way to a number of advanced bioimaging techniques applicable to living-cell studies such as sequential photolabeling of distinct cellular regions, innovative FRET schemes, or nanoscopy. Owing to the relevance of fluorescent proteins from Aequorea victoria (AFPs) for cell biology, a photochromic "toolbox" constituted by several AFPs is highly desirable. Here we introduce four new photochromic AFPs whose reversible photoswitching occurs between the native bright and a dark state at low illumination power, on account of a very efficient cis-trans photoisomerization. Most remarkably, the optical bistability of these AFPs derives from the single E222Q mutation in the primary sequence. Apparently, the E222Q substitution can restore the intrinsic photochromic behavior of the isolated chromophore. The significance of these mutants for high-resolution in vivo cell imaging is shown by means of photochromic FRET experiments. SN - 1520-5126 UR - https://www.unboundmedicine.com/medline/citation/19958004/Single_amino_acid_replacement_makes_Aequorea_victoria_fluorescent_proteins_reversibly_photoswitchable_ L2 - https://dx.doi.org/10.1021/ja9014953 DB - PRIME DP - Unbound Medicine ER -