RRC ID 47225
Author Ogoh K, Kinebuchi T, Murai M, Takahashi T, Ohmiya Y, Suzuki H.
Title Dual-color-emitting green fluorescent protein from the sea cactus Cavernularia obesa and its use as a pH indicator for fluorescence microscopy.
Journal Luminescence
Abstract We isolated and characterized a green fluorescent protein (GFP) from the sea cactus Cavernularia obesa. This GFP exists as a dimer and has absorption maxima at 388 and 498 nm. Excitation at 388 nm leads to blue fluorescence (456 nm maximum) at pH 5 and below, and green fluorescence (507 nm maximum) at pH 7 and above, and the GFP is remarkably stable at pH 4. Excitation at 498 nm leads to green fluorescence (507 nm maximum) from pH 5 to pH 9. We introduced five amino acid substitutions so that this GFP formed monomers rather than dimers and then used this monomeric form to visualize intracellular pH change during the phagocytosis of living cells by use of fluorescence microscopy. The intracellular pH change is visualized by use of a simple long-pass emission filter with single-wavelength excitation, which is technically easier to use than dual-emission fluorescent proteins that require dual-wavelength excitation.
Volume 28(4)
Pages 582-91
Published 2013-7-1
DOI 10.1002/bio.2497
PMID 23468077
PMC PMC3884763
MeSH Amino Acid Sequence Animals Anthozoa / chemistry* Cell Line Color* Green Fluorescent Proteins / chemistry* Green Fluorescent Proteins / genetics HeLa Cells Humans Hydrogen-Ion Concentration Indicators and Reagents / chemistry* Mice Microscopy, Fluorescence Molecular Sequence Data
IF 1.855
Times Cited 4
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
DNA material pCoGFP-wt (RDB14368) pCoGFP-mam (RDB14369) pCoGFP-V0 (RDB14370) pCoGFP-V1 (RDB14371) pCoGFP-V2 (RDB14372) pCoGFP-V3 (RDB14373) pCoGFP-V4 (RDB14374).