Identification of anti-TNFalpha peptides with consensus sequence.
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.
Zhang
Jie
J
Department of Biochemistry, Beijing Institute of Basic Medical Sciences, P.O. Box 130(3), Beijing 100850, People's Republic of China.
Zheng
Li
L
Zhao
An
A
Gao
Bo
B
Liu
Nong-Le
NL
Wang
Fang
F
Dong
Jie
J
Xin
Zhong-Tao
ZT
Shao
Ning-Sheng
NS
Wang
Hui-Xin
HX
Xue
Yan-Ning
YN
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
D017931:DNA Primers; D009838:Oligodeoxyribonucleotides; D010455:Peptides; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha