title: LanFP10-A, first functional fluorescent protein whose chromophore contains the elusive mutation G67A.
abstract: Since Green Fluorescent Protein (GFP) was first successfully expressed in heterologous systems in 1994, many genes encoding other natural autofluorescent proteins (AFPs) have been cloned and subsequently modified by protein engineering to improve their physicochemical properties. Throughout this twenty-two-year period, glycine 67 (Gly67) has been regarded as the only amino acid in the entire protein family that is essential for the formation of the different reported chromophores. In this work, we demonstrate that a synthetic gene encoding LanFP10-A, a natural protein encoded in the genome of the lancelet Branchiostoma floridae containing the G67A mutation, produces a heterologous, functional yellow fluorescent protein when expressed in E. coli. In contrast to LanFP10-A, LanFP6-A, a second GFP-like protein found in the lancelet genome that also contains the natural G67A mutation, was non-fluorescent.
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Col. Chamilpa, Cuernavaca, Morelos 62210, Mexico.
authors.forename: Abigail
authors.initials: A
authors.lastname: Roldán-Salgado
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Col. Chamilpa, Cuernavaca, Morelos 62210, Mexico.
authors.forename: Celidee
authors.initials: C
authors.lastname: Sánchez-Barreto
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Col. Chamilpa, Cuernavaca, Morelos 62210, Mexico. Electronic address: paul@ibt.unam.mx.
authors.forename: Paul
authors.initials: P
authors.lastname: Gaytán
mesh_terms: D000818:Animals; D005453:Fluorescence; D064175:Lancelets; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D020125:Mutation, Missense; D000072417:Protein Domains
chemical_list: D008164:Luminescent Proteins
keywords: Autofluorescent proteins; Branchiostoma; G67A mutation; GFP-like proteins; Genetic reporters; Protein engineering; Protein expression; Synthetic genes