title: Superoxide production in human neutrophils is enhanced by treatment with transmembrane peptides derived from human formyl peptide receptor.
abstract: Formyl peptide receptor (FPR) mediates a number of important host defense functions. Although studies have been performed on the ligand binding site of FPR, FPR dynamic behavior such as receptor dimerization on the cell surface remains unknown. Recently, peptides derived from the transmembrane (TM) domains of GPCRs were shown to disrupt dimer formation by receptors and to result in specific regulation of receptor function. To reveal the function of FPR TM domains, hFPRTM peptides derived from FPR were synthesized, and their biological activities were evaluated with human neutrophils. Synthetic peptides did not exhibit agonistic or antagonistic activity toward superoxide anion production. However, Neutrophils treated with hFPRTM4 produced 4-fold superoxide anion compared with untreated cells when stimulated with FPR agonist fMLP. Short peptide fragments derived from the fourth TM region of FPR did not enhance superoxide anion production, which suggests that hFPRTM4 did not behave as a ligand. CD and fluorescence spectra suggested that hFPRTM peptides were inserted into the membrane. The addition of hFPRTM4 increased the intracellular calcium concentration, which meant the peptide activated some membrane protein on the cell surface. The present study suggests that the fourth TM domain of FPR has a function related to a priming effect.
authors.affiliation: Department of Chemistry, Faculty of Science and Engineering, Saga University, Saga 840-8502.
authors.forename: Ryo
authors.initials: R
authors.lastname: Hayashi
authors.forename: Satoshi
authors.initials: S
authors.lastname: Osada
authors.forename: Masahiro
authors.initials: M
authors.lastname: Yoshiki
authors.forename: Daisuke
authors.initials: D
authors.lastname: Sugiyama
authors.forename: Ichiro
authors.initials: I
authors.lastname: Fujita
authors.forename: Yuhei
authors.initials: Y
authors.lastname: Hamasaki
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kodama
mesh_terms: D000595:Amino Acid Sequence; D002118:Calcium / Q000378:metabolism; D002942:Circular Dichroism; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism; D010455:Peptides / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology*; D044042:Receptors, Formyl Peptide / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013481:Superoxides / Q000378:metabolism*
chemical_list: D008565:Membrane Proteins; D010455:Peptides; D044042:Receptors, Formyl Peptide; D013481:Superoxides; D002118:Calcium