title: Peptides derived from CXCL8 based on in silico analysis inhibit CXCL8 interactions with its receptor CXCR1.
abstract: Chemokine CXCL8 is crucial for regulation of inflammatory and immune responses via activating its cognate receptor CXCR1. In this study, molecular docking and binding free energy calculations were combined to predict the initial binding event of CXCL8 to CXCR1 for peptide drug design. The simulations reveal that in the initial binding, the N-loop of CXCL8 interacts with the N-terminus of CXCR1, which is dominated by electrostatic interactions. The derived peptides from the binding region of CXCL8 are synthesized for further confirmation. Surface plasmon resonance analyses indicate that the CXCL8 derived peptide with 14 residues is able to bind to the receptor CXCR1 derived peptide with equilibrium KD of 252 μM while the peptide encompassing a CXCL8 K15A mutation hardly binds to CXCR1 derived peptide (KD = 1553 μM). The cell experiments show that the designed peptide inhibits CXCL8-induced and LPS-activated monocytes adhesion and transmigration. However, when the peptides were mutated on two lysine residues (K15 and K20), the inhibition effects were greatly reduced indicating these two amino acids are key residues for the initial binding of CXCL8 to CXCR1. This study demonstrates that in silico prediction based functional peptide design can be effective for developing anti-inflammation drugs.
authors.affiliation: Department of Biochemistry, School of Medicine, Tzu Chi University, Hualien 97004, Taiwan.
authors.forename: Shinn-Jong
authors.initials: SJ
authors.lastname: Jiang
authors.affiliation: Department of Biochemistry, School of Medicine, Tzu Chi University, Hualien 97004, Taiwan.|Institute of Medical Sciences, Tzu Chi University, Hualien 97004, Taiwan.
authors.forename: Je-Wen
authors.initials: JW
authors.lastname: Liou
authors.affiliation: Institute of Medical Sciences, Tzu Chi University, Hualien 97004, Taiwan.|Department of Laboratory Medicine, Tzu Chi Medical Center, Hualien 97004, Taiwan.
authors.forename: Chun-Chun
authors.initials: CC
authors.lastname: Chang
authors.affiliation: Department of Life Sciences, Tzu Chi University, Hualien 97004, Taiwan.
authors.forename: Yi
authors.initials: Y
authors.lastname: Chung
authors.affiliation: Department of Life Sciences, Tzu Chi University, Hualien 97004, Taiwan.
authors.forename: Lee-Fong
authors.initials: LF
authors.lastname: Lin
authors.affiliation: Department of Life Sciences, Tzu Chi University, Hualien 97004, Taiwan.
authors.forename: Hao-Jen
authors.initials: HJ
authors.lastname: Hsu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D003198:Computer Simulation; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects; D006801:Humans; D055767:Immobilized Proteins / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism*; D008024:Ligands; D008070:Lipopolysaccharides / Q000494:pharmacology; D062105:Molecular Docking Simulation; D008969:Molecular Sequence Data; D009000:Monocytes / Q000166:cytology / Q000187:drug effects; D050505:Mutant Proteins / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D011485:Protein Binding / Q000187:drug effects; D023062:Receptors, Interleukin-8A / Q000378:metabolism*; D015203:Reproducibility of Results; D020349:Surface Plasmon Resonance; D013499:Surface Properties; D013816:Thermodynamics
chemical_list: D055767:Immobilized Proteins; D016209:Interleukin-8; D008024:Ligands; D008070:Lipopolysaccharides; D050505:Mutant Proteins; D010455:Peptides; D023062:Receptors, Interleukin-8A