title: The first mutant of the Aequorea victoria green fluorescent protein that forms a red chromophore.
abstract: Green fluorescent protein (GFP) from a jellyfish, Aequorea victoria, and its mutants are widely used in biomedical studies as fluorescent markers. In spite of the enormous efforts of academia and industry toward generating its red fluorescent mutants, no GFP variants with emission maximum at more than 529 nm have been developed during the 15 years since its cloning. Here, we used a new strategy of molecular evolution aimed at generating a red-emitting mutant of GFP. As a result, we have succeeded in producing the first GFP mutant that substantially matures to the red-emitting state with excitation and emission maxima at 555 and 585 nm, respectively. A novel, nonoxidative mechanism for formation of the red chromophore in this mutant that includes a dehydration of the Ser65 side chain has been proposed. Model experiments showed that the novel dual-color GFP mutant with green and red emission is suitable for multicolor flow cytometry as an additional color since it is clearly separable from both green and red fluorescent tags.
authors: Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Miklukho-Maklaya 16/10, Moscow 117997, Russia.
authors: Alexander S
authors: AS
authors: Mishin
authors: Fedor V
authors: FV
authors: Subach
authors: Ilia V
authors: IV
authors: Yampolsky
authors: William
authors: W
authors: King
authors: Konstantin A
authors: KA
authors: Lukyanov
authors: Vladislav V
authors: VV
authors: Verkhusha
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003116:Color; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D037402:Hydrozoa / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: C485192:green fluorescent protein, Aequorea victoria; D049452:Green Fluorescent Proteins