title: G196 epitope tag system: a novel monoclonal antibody, G196, recognizes the small, soluble peptide DLVPR with high affinity.
abstract: The recognition specificity of monoclonal antibodies (mAbs) has made mAbs among the most frequently used tools in both basic science research and in clinical diagnosis and therapies. Precise determination of the epitope allows the development of epitope tag systems to be used with recombinant proteins for various purposes. Here we describe a new family of tag derived from the epitope recognized by a highly specific mAb G196. The minimal epitope was identified as the five amino acid sequence Asp-Leu-Val-Pro-Arg. Permutation analysis was used to characterize the binding requirements of mAb G196, and the variable regions of the mAb G196 were identified and structurally analyzed by X-ray crystallography. Isothermal titration calorimetry revealed the high affinity (Kd= 1.25 nM) of the mAb G196/G196-epitope peptide interaction, and G196-tag was used to detect several recombinant cytosolic and nuclear proteins in human and yeast cells. mAb G196 is valuable for developing a new peptide tagging system for cell biology and biochemistry research.
authors.affiliation: Department of Biochemistry, Shimane University School of Medicine, Izumo 693-8501, Japan.|Department of Oral and Maxillofacial Surgery Shimane University School of Medicine, Izumo 693-8501, Japan.
authors.forename: Kasumi
authors.initials: K
authors.lastname: Tatsumi
authors.affiliation: Department of Biochemistry, Shimane University School of Medicine, Izumo 693-8501, Japan.
authors.forename: Gyosuke
authors.initials: G
authors.lastname: Sakashita
authors.affiliation: Department of Biochemistry, Shimane University School of Medicine, Izumo 693-8501, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Nariai
authors.affiliation: Department of Biochemistry, Shimane University School of Medicine, Izumo 693-8501, Japan.
authors.forename: Kosuke
authors.initials: K
authors.lastname: Okazaki
authors.affiliation: Department of Biochemistry, Shimane University School of Medicine, Izumo 693-8501, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kato
authors.affiliation: Department of Biochemistry, Shimane University School of Medicine, Izumo 693-8501, Japan.
authors.forename: Eiji
authors.initials: E
authors.lastname: Obayashi
authors.affiliation: Drug Design Group, Kanagawa Academy of Science and Technology, Kawasaki, Kanagawa, 213-0012, Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Yoshida
authors.affiliation: Drug Design Group, Kanagawa Academy of Science and Technology, Kawasaki, Kanagawa, 213-0012, Japan.
authors.forename: Kanako
authors.initials: K
authors.lastname: Sugiyama
authors.affiliation: Drug Design Group, Kanagawa Academy of Science and Technology, Kawasaki, Kanagawa, 213-0012, Japan.|Protein Design Laboratory, Graduate School of Medical Life Science, Yokohama City University, Tsurumi, Yokohama 230-0045, Japan.
authors.forename: Sam-Yong
authors.initials: SY
authors.lastname: Park
authors.affiliation: Department of Oral and Maxillofacial Surgery Shimane University School of Medicine, Izumo 693-8501, Japan.
authors.forename: Joji
authors.initials: J
authors.lastname: Sekine
authors.affiliation: Department of Biochemistry, Shimane University School of Medicine, Izumo 693-8501, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Urano
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D000915:Antibody Affinity; D000918:Antibody Specificity; D001665:Binding Sites; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D018604:Epitope Mapping / Q000379:methods*; D000939:Epitopes / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D011485:Protein Binding; D000072756:Protein Conformation, alpha-Helical; D000072757:Protein Conformation, beta-Strand; D054730:Protein Interaction Domains and Motifs; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D010455:Peptides; D011994:Recombinant Proteins