Tags

Type your tag names separated by a space and hit enter

Conversion of the monomeric red fluorescent protein into a photoactivatable probe.
Chem Biol. 2005 Mar; 12(3):279-85.CB

Abstract

Photoactivatable fluorescent proteins bring new dimension to the analysis of protein dynamics in the cell. Protein tagged with a photoactivatable label can be visualized and tracked in a spatially and temporally defined manner. Here, we describe a basic rational design strategy to develop monomeric photoactivatable proteins using site-specific mutagenesis of common monomeric red-shifted fluorescent proteins. This strategy was applied to mRFP1, which was converted into probes that are photoactivated by either green or violet light. The latter photoactivatable variants, named PA-mRFP1s, exhibited a 70-fold increase of fluorescence intensity resulting from the photoconversion of a violet-light-absorbing precursor. Detailed characterization of PA-mRFP1s was performed with the purified proteins and the proteins expressed in mammalian cells where the photoactivatable properties were preserved. PA-mRFP1s were used as protein tags to study the intracellular dynamics of GTPase Rab5.

Authors+Show Affiliations

Department of Pharmacology, University of Colorado Health Sciences Center, Aurora, CO 80045, USA. vladislav.verkhusha@uchsc.eduNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

15797211

Citation

Verkhusha, Vladislav V., and Alexander Sorkin. "Conversion of the Monomeric Red Fluorescent Protein Into a Photoactivatable Probe." Chemistry & Biology, vol. 12, no. 3, 2005, pp. 279-85.
Verkhusha VV, Sorkin A. Conversion of the monomeric red fluorescent protein into a photoactivatable probe. Chem Biol. 2005;12(3):279-85.
Verkhusha, V. V., & Sorkin, A. (2005). Conversion of the monomeric red fluorescent protein into a photoactivatable probe. Chemistry & Biology, 12(3), 279-85.
Verkhusha VV, Sorkin A. Conversion of the Monomeric Red Fluorescent Protein Into a Photoactivatable Probe. Chem Biol. 2005;12(3):279-85. PubMed PMID: 15797211.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Conversion of the monomeric red fluorescent protein into a photoactivatable probe. AU - Verkhusha,Vladislav V, AU - Sorkin,Alexander, PY - 2004/10/01/received PY - 2005/01/07/revised PY - 2005/01/10/accepted PY - 2005/3/31/pubmed PY - 2005/8/10/medline PY - 2005/3/31/entrez SP - 279 EP - 85 JF - Chemistry & biology JO - Chem. Biol. VL - 12 IS - 3 N2 - Photoactivatable fluorescent proteins bring new dimension to the analysis of protein dynamics in the cell. Protein tagged with a photoactivatable label can be visualized and tracked in a spatially and temporally defined manner. Here, we describe a basic rational design strategy to develop monomeric photoactivatable proteins using site-specific mutagenesis of common monomeric red-shifted fluorescent proteins. This strategy was applied to mRFP1, which was converted into probes that are photoactivated by either green or violet light. The latter photoactivatable variants, named PA-mRFP1s, exhibited a 70-fold increase of fluorescence intensity resulting from the photoconversion of a violet-light-absorbing precursor. Detailed characterization of PA-mRFP1s was performed with the purified proteins and the proteins expressed in mammalian cells where the photoactivatable properties were preserved. PA-mRFP1s were used as protein tags to study the intracellular dynamics of GTPase Rab5. SN - 1074-5521 UR - https://www.unboundmedicine.com/medline/citation/15797211/Conversion_of_the_monomeric_red_fluorescent_protein_into_a_photoactivatable_probe_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1074-5521(05)00026-8 DB - PRIME DP - Unbound Medicine ER -