title: Selection of CC chemokine receptor 5-binding peptide from a phage display peptide library.
abstract: Human CC chemokine receptor (CCR) 5 is a G protein-coupled receptor involved in a broad range of human diseases that mediates HIV-1 viral entry into cells. Certain small molecule receptor antagonists to CCR5 have been useful in therapy for these diseases. In this study, CCR5-expressing CHO cells (CHO/CCR5 cells) were used to select CCR5-binding peptides from a phage-displayed 12-mers peptide library. All of the 30 clones selected from the library showed specific binding to CHO/CCR5 cells by enzyme linked immunosorbent assay (ELISA). Seventeen out of the 30 clones shared the amino acid motif AFDWTFVPSLIL. The motif-containing phages and synthetic peptide AFDWTFVPSLIL blocked the binding of mAb 2D7 to CHO/CCR5 cells and competitively inhibited the ability of chemokine regulated on activation normal T cell expressed and secreted (RANTES) binding to CHO/CCR5 cells. Furthermore, the peptide AFDWTFVPSLIL also inhibited RANTES induced increase in the intracellular Ca2+ level in CHO/CCR5 cells. These results suggest that the peptide AFDWTFVPSLIL was specific for CCR5 and that it might become a CCR5 antagonist.
authors.affiliation: Department of Biochemistry, The Key Laboratory of Genetic Engineering of the Ministry of Education, Sun Yat-sen University, Guanzhou, P R China.
authors.forename: Fang-Yu
authors.initials: FY
authors.lastname: Wang
authors.forename: Tian-Yuan
authors.initials: TY
authors.lastname: Zhang
authors.forename: Jin-Xian
authors.initials: JX
authors.lastname: Luo
authors.forename: Guo-An
authors.initials: GA
authors.lastname: He
authors.forename: Qu-Liang
authors.initials: QL
authors.lastname: Gu
authors.forename: Fan
authors.initials: F
authors.lastname: Xiao
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001483:Base Sequence; D001667:Binding, Competitive; D065100:CCR5 Receptor Antagonists; D016466:CHO Cells; D020013:Calcium Signaling; D018946:Chemokine CCL5 / Q000378:metabolism; D006224:Cricetinae; D006801:Humans; D019151:Peptide Library; D011485:Protein Binding; D019713:Receptors, CCR5 / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D065100:CCR5 Receptor Antagonists; D018946:Chemokine CCL5; D019151:Peptide Library; D019713:Receptors, CCR5