title: Identification of cell binding sites in the laminin alpha5-chain G domain.
abstract: The laminins consist of at least 11 polypeptides (5 alpha-chains, 3 beta-chains, and 3 gamma-chains) specific to basement membranes. Here we investigate the biological activity associated with the G domain of the newly identified laminin alpha5-chain using 113 overlapping synthetic peptides (positions 2679-3635). Using HT-1080 cells, 21 peptides showed attachment activity either on peptide-coated tissue culture plates or to peptide-conjugated Sepharose beads. Heparin inhibited cell attachment to 16 peptides, while ethylenediaminetetraacetic acid exhibited no inhibitory activity. Peptides A5G-27, A5G-65, and A5G-71 showed the strongest cell attachment, with the minimum active core sequences of the peptides being GIIFFL, HQNMGSVNVSV, and YLQFVG, respectively. Furthermore, these 16 peptides were tested for their ability to stimulate neurite outgrowth in the PC12 cells. A5G-3, A5G-33, A5G-71, A5G-73, A5G-81, and A5G-101 were the only peptides of the 16 that demonstrated the ability to promote neurite outgrowth. These results demonstrate that synthetic peptides with alpha5-chain G domain primary amino acid sequences possess some of the same biological activities attributable to the whole laminin and the alpha5-chain G domain. Therefore, these peptides may be useful in the investigation of laminin-receptor interactions and possibly mechanisms of laminin signal transduction.
authors.affiliation: Graduate School of Environmental Earth Science, Hokkaido University, Kita 10 Nishi 5, Kita-ku, Sapporo 060-0810, Japan.
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Makino
authors.forename: Ikuko
authors.initials: I
authors.lastname: Okazaki
authors.forename: Shingo
authors.initials: S
authors.lastname: Kasai
authors.forename: Norio
authors.initials: N
authors.lastname: Nishi
authors.forename: Maria
authors.initials: M
authors.lastname: Bougaeva
authors.forename: Benjamin S
authors.initials: BS
authors.lastname: Weeks
authors.forename: Akira
authors.initials: A
authors.lastname: Otaka
authors.forename: Peter K
authors.initials: PK
authors.lastname: Nielsen
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Yamada
authors.forename: Motoyoshi
authors.initials: M
authors.lastname: Nomizu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002455:Cell Division / Q000187:drug effects; D006801:Humans; D007797:Laminin / Q000138:chemical synthesis / Q000378:metabolism*; D008969:Molecular Sequence Data; D016501:Neurites / Q000187:drug effects / Q000502:physiology; D016716:PC12 Cells; D010455:Peptides / Q000138:chemical synthesis / Q000378:metabolism*; D010969:Plastics; D017434:Protein Structure, Tertiary; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012685:Sepharose; D014407:Tumor Cells, Cultured
chemical_list: D007797:Laminin; D010455:Peptides; D010969:Plastics; D011993:Recombinant Fusion Proteins; C119013:laminin 10; C097043:laminin alpha5; D012685:Sepharose