title: Identification of anti-TNFalpha peptides with consensus sequence.
abstract: Phage displayed peptide library was used to select tumor necrosis factor alpha (TNFalpha) binding peptides. After three sequential rounds of biopanning, some linear TNFalpha-binding peptides were identified from a 12-mer peptide library. A consensus sequence (L/M)HEL(Y/F)(L/M)X(W/Y/F), where X might be variable residue, was deduced from sequences of these peptides. The phages bearing these peptides showed specific binding to immobilized TNFalpha, with over 80% of phages bound being competitively eluted by free TNFalpha. To confirm the binding activity and to explore further functional properties, three peptides with typical structure were selected and expressed as GST-fused protein. These recombinant peptides effectively competed for [125I]TNFalpha binding to TNFR1 in a dose-dependent manner, with IC(50) from 10 to 160 microM. Furthermore, the GST-fused derivatives showed inhibitory effects on TNFalpha-induced cytotoxicity. Taken together, these data demonstrate that the TNFalpha-binding peptides are effective antagonists of TNFalpha and the deduced motif might be useful in development of novel low molecular weight anti-TNFalpha drugs.
authors.affiliation: Department of Biochemistry, Beijing Institute of Basic Medical Sciences, P.O. Box 130(3), Beijing 100850, People's Republic of China.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Li
authors.initials: L
authors.lastname: Zheng
authors.forename: An
authors.initials: A
authors.lastname: Zhao
authors.forename: Bo
authors.initials: B
authors.lastname: Gao
authors.forename: Nong-Le
authors.initials: NL
authors.lastname: Liu
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.forename: Jie
authors.initials: J
authors.lastname: Dong
authors.forename: Zhong-Tao
authors.initials: ZT
authors.lastname: Xin
authors.forename: Ning-Sheng
authors.initials: NS
authors.lastname: Shao
authors.forename: Hui-Xin
authors.initials: HX
authors.lastname: Wang
authors.forename: Yan-Ning
authors.initials: YN
authors.lastname: Xue
mesh_terms: D001435:Bacteriophages / Q000737:chemistry; D001483:Base Sequence; D016384:Consensus Sequence; D017931:DNA Primers; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010455:Peptides / Q000737:chemistry*; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000737:chemistry
chemical_list: D017931:DNA Primers; D009838:Oligodeoxyribonucleotides; D010455:Peptides; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha