title: Identification of novel peptides against TNF-α using phage display technique and in silico modeling of their modes of binding.
abstract: The aim of this study was to identify novel TNF-α blocking peptide(s) using phage display technology. Two novel 7-mer TNF-α binding peptides P51 and P52 with Kdvalues of 1.47 and 0.51nM were identified. Phage particles displaying P51 and P52 peptides at 0.318nM concentration prevent cytotoxic effect of TNF-α on L929 cells by 8.2% and 16.15%, respectively. Synthesized P51 and P52 peptides also inhibited TNF-α induced cytotoxicity with IC50values of 25.15±2.18 and 7.08±2.24μM, respectively. The result of RT-PCR also supports the inhibitory activity of the identified peptides, where P51 and P52 significantly inhibit the inductive effect of TNF-α on IκB-α mRNA levels. The inhibitory effects of the peptides were attributed to their abilities of binding at the inter-subunit interfaces leading to TNF-α dissociation. The results of molecular docking studies revealed that the peptides-TNF-α complexes are mostly stabilized by hydrophobic contacts.
authors.affiliation: Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; School of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Ali Akbar
authors.initials: AA
authors.lastname: Alizadeh
authors.affiliation: Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; School of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran. Electronic address: hamzehm@tbzmed.ac.ir.
authors.forename: Maryam
authors.initials: M
authors.lastname: Hamzeh-Mivehroud
authors.affiliation: Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Malak
authors.initials: M
authors.lastname: Farajzadeh
authors.affiliation: Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; School of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran. Electronic address: dastmalchi.s@tbzmed.ac.ir.
authors.forename: Siavoush
authors.initials: S
authors.lastname: Dastmalchi
mesh_terms: D000818:Animals; D061505:Cell Surface Display Techniques / Q000379:methods*; D003198:Computer Simulation; D006801:Humans; D019169:Jurkat Cells; D051379:Mice; D008956:Models, Chemical; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*
chemical_list: D019151:Peptide Library; D010455:Peptides; D014409:Tumor Necrosis Factor-alpha
keywords: MTT assay; Molecular docking; Peptide ligand; Phage display; RT-PCR; TNF-α