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A colourless green fluorescent protein homologue from the non-fluorescent hydromedusa Aequorea coerulescens and its fluorescent mutants.
Biochem J. 2003 Jul 15; 373(Pt 2):403-8.BJ

Abstract

We have cloned an unusual colourless green fluorescent protein (GFP)-like protein from Aequorea coerulescens (acGFPL). The A. coerulescens specimens displayed blue (not green) luminescence, and no fluorescence was detected in these medusae. Escherichia coli expressing wild-type acGFPL showed neither fluorescence nor visible coloration. Random mutagenesis generated green fluorescent mutants of acGFPL, with the strongest emitters found to contain an Glu(222)-->Gly (E222G) substitution, which removed the evolutionarily invariant Glu(222). Re-introduction of Glu(222) into the most fluorescent random mutant, named aceGFP, converted it into a colourless protein. This colourless aceGFP-G222E protein demonstrated a novel type of UV-induced photoconversion, from an immature non-fluorescent form into a green fluorescent form. Fluorescent aceGFP may be a useful biological tool, as it was able to be expressed in a number of mammalian cell lines. Furthermore, expression of a fusion protein of 'humanized' aceGFP and beta-actin produced a fluorescent pattern consistent with actin distribution in mammalian cells.

Authors+Show Affiliations

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, Moscow 117997, Russia.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

12693991

Citation

Gurskaya, Nadya G., et al. "A Colourless Green Fluorescent Protein Homologue From the Non-fluorescent Hydromedusa Aequorea Coerulescens and Its Fluorescent Mutants." The Biochemical Journal, vol. 373, no. Pt 2, 2003, pp. 403-8.
Gurskaya NG, Fradkov AF, Pounkova NI, et al. A colourless green fluorescent protein homologue from the non-fluorescent hydromedusa Aequorea coerulescens and its fluorescent mutants. Biochem J. 2003;373(Pt 2):403-8.
Gurskaya, N. G., Fradkov, A. F., Pounkova, N. I., Staroverov, D. B., Bulina, M. E., Yanushevich, Y. G., Labas, Y. A., Lukyanov, S., & Lukyanov, K. A. (2003). A colourless green fluorescent protein homologue from the non-fluorescent hydromedusa Aequorea coerulescens and its fluorescent mutants. The Biochemical Journal, 373(Pt 2), 403-8.
Gurskaya NG, et al. A Colourless Green Fluorescent Protein Homologue From the Non-fluorescent Hydromedusa Aequorea Coerulescens and Its Fluorescent Mutants. Biochem J. 2003 Jul 15;373(Pt 2):403-8. PubMed PMID: 12693991.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A colourless green fluorescent protein homologue from the non-fluorescent hydromedusa Aequorea coerulescens and its fluorescent mutants. AU - Gurskaya,Nadya G, AU - Fradkov,Arkady F, AU - Pounkova,Natalia I, AU - Staroverov,Dmitry B, AU - Bulina,Maria E, AU - Yanushevich,Yurii G, AU - Labas,Yulii A, AU - Lukyanov,Sergey, AU - Lukyanov,Konstantin A, PY - 2003/04/14/accepted PY - 2003/03/04/revised PY - 2002/12/19/received PY - 2003/4/16/pubmed PY - 2003/8/16/medline PY - 2003/4/16/entrez SP - 403 EP - 8 JF - The Biochemical journal JO - Biochem. J. VL - 373 IS - Pt 2 N2 - We have cloned an unusual colourless green fluorescent protein (GFP)-like protein from Aequorea coerulescens (acGFPL). The A. coerulescens specimens displayed blue (not green) luminescence, and no fluorescence was detected in these medusae. Escherichia coli expressing wild-type acGFPL showed neither fluorescence nor visible coloration. Random mutagenesis generated green fluorescent mutants of acGFPL, with the strongest emitters found to contain an Glu(222)-->Gly (E222G) substitution, which removed the evolutionarily invariant Glu(222). Re-introduction of Glu(222) into the most fluorescent random mutant, named aceGFP, converted it into a colourless protein. This colourless aceGFP-G222E protein demonstrated a novel type of UV-induced photoconversion, from an immature non-fluorescent form into a green fluorescent form. Fluorescent aceGFP may be a useful biological tool, as it was able to be expressed in a number of mammalian cell lines. Furthermore, expression of a fusion protein of 'humanized' aceGFP and beta-actin produced a fluorescent pattern consistent with actin distribution in mammalian cells. SN - 0264-6021 UR - https://www.unboundmedicine.com/medline/citation/12693991/A_colourless_green_fluorescent_protein_homologue_from_the_non_fluorescent_hydromedusa_Aequorea_coerulescens_and_its_fluorescent_mutants_ L2 - https://portlandpress.com/biochemj/article-lookup/doi/10.1042/BJ20021966 DB - PRIME DP - Unbound Medicine ER -