Bright photoconvertible fluorescent protein with high sensitivity
- Photoconvertible fluorescent protein
- Monomer/tetramer
- Useful for labeling proteins and subcellular structures
- High-sensitivity and brightness
The fluorescent protein Kikume Green gene was isolated from the stony coral Favia favus (Kikume-ish in Japanese).
CoralHue® Kikume Green-Red1 (KikGR1) protein emits bright green fluorescence that can be irreversibly converted to red. KikGR1 can be photoconverted by irradiation with UV or violet light (350-410 nm). Photoconverted KikGR1 possesses excitation/emission maxima at 583/593 nm. KikGR1 contains a His62-Tyr63-Gly64 tripeptide sequence which forms a green chromophore that can be photoconverted to a red one via formal beta-elimination and subsequent extension of the π-conjugated system. CoralHue® monomeric Kikume Green-Red (mKikGR1) maintains the brightness of the parent protein KikGR1. mKikGR1 can be used for labeling proteins or subcellular structures.
Images are provided by Dr.Tsutsui Hidekazu and Dr. Miyawaki Atsushi, Laboratory for Cell Function and Dynamics, BSI, RIKEN
Performance and use
KikGR1 transgenic zebrafish
The KikGR1 transgenic zebrafish was observed using two-photon excitation microscopy. KikGR1 can be photoconverted using two-photon excitation microscopy with the laser tuned to 780 nm.
Cell tracking using a photoconvertible fluorescent protein.Hatta K, Tsujii H, Omura T.
Nat Protoc. (2006) 1: 960-967.
Citations
- Tsutsui H, Karasawa S, Shimizu H, Nukina N, Miyawaki A. Semi-rational engineering of a coral fluorescent protein into an efficient highlighter. EMBO Rep. (2005) 6: 233-238 . PMID: 15731765
- Hatta K, Tsujii H, Omura T. Cell tracking using a photoconvertible fluorescent protein. Nat Protoc. (2006) : 960-967. PMID: 17406330
- Habuchi S, Tsutsui H, Kochaniak AB, Miyawaki A, van Oijen AM. van Oijen mKikGR, a Monomeric Photoswitchable Fluorescent Protein. PLoS ONE (2008) 3: e3944 PMID: 19079591
- Tsutsui H, Shimizu H, Mizuno H, Nukina N, Furuta T, Miyawaki A. The E1 mechanism in photo-induced beta-elimination reactions for green-to-red conversion of fluorescent proteins. Chem Biol. (2009) 16:1140-7. PMID: 19942137
- Mizuno H, Dedecker P, Ando R, Fukano T, Hofkens J, Miyawaki A Higher resolution in localization microscopy by slower switching of a photochromic protein. Photochem Photobiol Sci. (2010) 2: 239-248. PMID: 20126801
Fluorescent properties
Kikume Green-Red(KikGR) spectrum data files (text files)
- mKikume Green excitation (4K)
- mKikume Green emission (4K)
- mKikume Red excitation (4K)
- mKikume Red emission (4K)
- Kikume Green excitation (4K)
- Kikume Green emission (4K)
- Kikume Red excitaton (4K)
- Kikume Red emission (4K)
Note: The file is in a tab-delimited text format. It contains values of the wavelength (1nm spacing) and brightness (fluorescence intensity peak value normalized to 1). Use a spreadsheet program to create a spectrum that will help you in choosing the appropriate excitation filter, dichroic mirror and fluorescence filter.
Recommended antibodies
CoralHue®Kikume Green-Red can be recognized using antibodies as shown below.
- Anti-Kikume Green-Red fore WB, Code No. M128-3M
WB: Western blotting, IP: Immunoprecipitation