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
IF 4.238
Times Cited 122
Resource
DNA material AG/pRSETB (RDB15228) hmAG407/pRSETB (RDB15229) hmAG1/pRSETB (RDB15230) CoralHue™ Azami-Green (pAG-S1) (RDB18244) CoralHue™ humanized monomeric Azami-Green 1 (phmAG1-MNL) (RDB18245) CoralHue™ monomeric Azami-Green 1 (pmAG1-S1) (RDB18246) CoralHue™ monomeric Azami-Green 1 (pmAG1-MC1) (RDB18247) CoralHue™ monomeric Azami-Green 1 (pmAG1-MN1) (RDB18248) CoralHue™ humanized monomeric Azami-Green 1 (phmAG1-S1) (RDB18249) CoralHue™ humanized monomeric Azami-Green 1 (phmAG1-MC1) (RDB18250) CoralHue™ humanized monomeric Azami-Green 1 (phmAG1-MN1) (RDB18251) CoralHue™ humanized monomeric Azami-Green 1 (phmAG1-MCL) (RDB18252) CoralHue™ Mitochondria-targeted monomeric Azami-Green (pMT-mAG1) (RDB18311) CoralHue™ ER-targeted monomeric Azami-Green 1 (pER-mAG1) (RDB18312) CoralHue™ Plasma Membrane-targeted monomeric Azami-Green 1 (pPM-mAG1) (RDB18313) CoralHue™ β-Actin-targeted monomeric Azami-Green 1 (pActin-mAG1) (RDB18314) CoralHue™ Nucleoplasm-targeted Azami-Green (pNP-AG) (RDB18315) CoralHue™ ER-targeted monomeric Azami-Green407 (pER-mAG407) (RDB18326) CoralHue™ humanized monomeric Azami-Green407 (phmAG407-S1) (RDB18328)