RRC ID 35468
Author Kumagai A, Ando R, Miyatake H, Greimel P, Kobayashi T, Hirabayashi Y, Shimogori T, Miyawaki A.
Title A bilirubin-inducible fluorescent protein from eel muscle.
Journal Cell
Abstract The fluorescent protein toolbox has revolutionized experimental biology. Despite this advance, no fluorescent proteins have been identified from vertebrates, nor has chromogenic ligand-inducible activation or clinical utility been demonstrated. Here, we report the cloning and characterization of UnaG, a fluorescent protein from Japanese eel. UnaG belongs to the fatty-acid-binding protein (FABP) family, and expression in eel is restricted to small-diameter muscle fibers. On heterologous expression in cell lines or mouse brain, UnaG produces oxygen-independent green fluorescence. Remarkably, UnaG fluorescence is triggered by an endogenous ligand, bilirubin, a membrane-permeable heme metabolite and clinical health biomarker. The holoUnaG structure at 1.2 Å revealed a biplanar coordination of bilirubin by reversible π-conjugation, and we used this high-affinity and high-specificity interaction to establish a fluorescence-based human bilirubin assay with promising clinical utility. UnaG will be the prototype for a versatile class of ligand-activated fluorescent proteins, with applications in research, medicine, and bioengineering.
Volume 153(7)
Pages 1602-11
Published 2013-6-20
DOI 10.1016/j.cell.2013.05.038
PII S0092-8674(13)00644-2
PMID 23768684
MeSH Amino Acid Sequence Animals Bilirubin / metabolism Cloning, Molecular Eels / metabolism* Fish Proteins / chemistry Fish Proteins / genetics* Fish Proteins / isolation & purification* Fish Proteins / metabolism Green Fluorescent Proteins / chemistry Green Fluorescent Proteins / genetics Green Fluorescent Proteins / isolation & purification* Green Fluorescent Proteins / metabolism Molecular Sequence Data Muscles / metabolism Sequence Alignment
IF 31.398
Times Cited 62
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY CELL BIOLOGY
Resource
DNA material UnaG/pcDNA3-FLAG (RDB15703) UnaG/pGEX-2T (RDB15704) pET-His UNAG (RDB13504)