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A green-emitting fluorescent protein from Galaxeidae coral and its monomeric version for use in fluorescent labeling.
J Biol Chem. 2003 Sep 05; 278(36):34167-71.JB

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.

Authors+Show Affiliations

Laboratory for Cell Function and Dynamics, Advanced Technology Development Group, Brain Science Institute, RIKEN 2-1 Hirosawa, Wako-City, Saitama 351-0198, Japan.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12819206

Citation

Karasawa, Satoshi, et al. "A Green-emitting Fluorescent Protein From Galaxeidae Coral and Its Monomeric Version for Use in Fluorescent Labeling." The Journal of Biological Chemistry, vol. 278, no. 36, 2003, pp. 34167-71.
Karasawa S, Araki T, Yamamoto-Hino M, et al. A green-emitting fluorescent protein from Galaxeidae coral and its monomeric version for use in fluorescent labeling. J Biol Chem. 2003;278(36):34167-71.
Karasawa, S., Araki, T., Yamamoto-Hino, M., & Miyawaki, A. (2003). A green-emitting fluorescent protein from Galaxeidae coral and its monomeric version for use in fluorescent labeling. The Journal of Biological Chemistry, 278(36), 34167-71.
Karasawa S, et al. A Green-emitting Fluorescent Protein From Galaxeidae Coral and Its Monomeric Version for Use in Fluorescent Labeling. J Biol Chem. 2003 Sep 5;278(36):34167-71. PubMed PMID: 12819206.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A green-emitting fluorescent protein from Galaxeidae coral and its monomeric version for use in fluorescent labeling. AU - Karasawa,Satoshi, AU - Araki,Toshio, AU - Yamamoto-Hino,Miki, AU - Miyawaki,Atsushi, Y1 - 2003/06/19/ PY - 2003/6/24/pubmed PY - 2003/10/8/medline PY - 2003/6/24/entrez SP - 34167 EP - 71 JF - The Journal of biological chemistry JO - J. Biol. Chem. VL - 278 IS - 36 N2 - 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. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/12819206/A_green_emitting_fluorescent_protein_from_Galaxeidae_coral_and_its_monomeric_version_for_use_in_fluorescent_labeling_ L2 - http://www.jbc.org/cgi/pmidlookup?view=long&pmid=12819206 DB - PRIME DP - Unbound Medicine ER -