RRC ID 50516
Author Karasawa S, Araki T, Yamamoto-Hino M, Miyawaki A.
Title A green-emitting fluorescent protein from Galaxeidae coral and its monomeric version for use in fluorescent labeling.
Journal J. Biol. Chem.
Abstract We have cloned a gene which encodes a fluorescent protein from the stony coral, Galaxeidae. This protein absorbs light maximally at 492 nm and emits green light at 505 nm, and as a result, we have designated it "Azami-Green (AG)." Despite sharing a similar spectral profile with enhanced green fluorescent protein (EGFP) (Clontech), the most popular variant of the Aequorea victoria green fluorescent protein, the identity between these two proteins at the amino acid level is only 5.7%. However, since AG has a high extinction coefficient, fluorescence quantum yield, and acid stability, it produces brighter green fluorescence in cultured cells than EGFP. Similar to other fluorescent proteins isolated from coral animals, AG forms a tight tetrameric complex, resulting in poor labeling of subcellular structures such as the plasma membrane and mitochondria. We have converted tetrameric AG into a monomeric form by the introduction of three amino acid substitutions, which were recently reported to be effective for monomerizing the red fluorescent protein from Discosoma coral (DsRed, Clontech). The resultant monomeric AG allowed for efficient fluorescent labeling of all of the subcellular structures and proteins tested while retaining nearly all of the brightness of the original tetrameric form. Thus, monomeric AG is a useful monomeric green-emitting fluorescent protein comparable to EGFP.
Volume 278(36)
Pages 34167-71
Published 2003-9-5
DOI 10.1074/jbc.M304063200
PII M304063200
PMID 12819206
MeSH Amino Acid Sequence Animals Anthozoa / metabolism* Cell Membrane / metabolism Chromatography Cloning, Molecular Cytosol / metabolism DNA, Complementary / metabolism Dogs Electrophoresis, Polyacrylamide Gel Green Fluorescent Proteins HeLa Cells Humans Hydrogen-Ion Concentration Light Luminescent Proteins / chemistry* Luminescent Proteins / metabolism* Luminescent Proteins / pharmacology* Mitochondria / metabolism Molecular Sequence Data Protein Transport Scattering, Radiation Sequence Homology, Amino Acid Spectrophotometry Time Factors Transfection
IF 4.011
Resource
DNA material AG/pRSETB (RDB15228) hmAG407/pRSETB (RDB15229) hmAG1/pRSETB (RDB15230).