title: A luminescent affinity tag for proteins based on the terbium(III)-binding peptide.
abstract: Genetically encoded tags attached to proteins of interest are widely exploited for proteome analysis. Here, we present Tb(3+)-binding peptides (TBPs) which can be used for both luminescent measurements and affinity purification of proteins. TBPs consist of acidic amino acid residues and tryptophan residues which serve as Tb(3+)-binding sites and sensitizers for Tb(3+) luminescence, respectively. The Tb(3+) complexes of TBPs fused to a target protein exhibited luminescence characteristic of Tb(3+) by excitation of the tryptophan residue, and fusion proteins fused to one of the TPBs were successfully isolated from Escherichia coli cell lysate by affinity chromatography with a Tb(3+)-immobilized solid support.
authors.affiliation: Department of Bioscience and Bioinformatics, Kyushu Institute of Technology, Iizuka, Japan. sueda@bio.kyutech.ac.jp
authors.forename: Shinji
authors.initials: S
authors.lastname: Sueda
authors.forename: Shogo
authors.initials: S
authors.lastname: Tanaka
authors.forename: Sayomi
authors.initials: S
authors.lastname: Inoue
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Komatsu
mesh_terms: D000345:Affinity Labels / Q000737:chemistry*; D000595:Amino Acid Sequence; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry*; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000737:chemistry; D005982:Glutathione Transferase / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000737:chemistry; D049408:Luminescent Agents / Q000737:chemistry; D008163:Luminescent Measurements / Q000379:methods*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D013725:Terbium / Q000737:chemistry*; D014364:Tryptophan / Q000737:chemistry
chemical_list: D000345:Affinity Labels; D002352:Carrier Proteins; D049408:Luminescent Agents; D010455:Peptides; C485184:enhanced green fluorescent protein; D013725:Terbium; D049452:Green Fluorescent Proteins; D014364:Tryptophan; D005982:Glutathione Transferase