title: Cyclic peptide analysis of the biologically active loop region in the laminin alpha3 chain LG4 module demonstrates the importance of peptide conformation on biological activity.
abstract: The laminin alpha3 chain LG4 module (alpha3LG4 module) has cell adhesion, heparin binding, migration, and neurite outgrowth activities. The LG4 module consists of a 14-stranded beta-sheet (A-N) sandwich structure. Previously, we identified the A3G756 sequence (KNSFMALYLSKGRLVFALG in the human laminin alpha3 chain 1411-1429) as a biologically active site in the alpha3LG4 module. The A3G756 sequence is located on the E and F strands based on a crystal structure-based sequence alignment. The Lys1421 and Arg1423 residues, critical amino acids for the biological activity of A3G756, are located on the E-F connecting loop region as a KGR sequence. In this study, we focused on the KGR sequence and investigated the structural requirements of the E-F connecting loop region in the alpha3LG4 module. We synthesized three linear peptides containing the KGR sequence at the middle and the N and C termini and also prepared three cyclic analogues corresponding to the linear peptides. cyclo-hEF3A (CLYLSKGRLVFAC), which is a cyclic peptide containing the KGR sequence at the middle, showed the strongest inhibitory effect on both the heparin binding and the cell attachment to the recombinant alpha3LG4 module protein. The cyclo-hEF3A peptide was more active for syndecan-4 binding and neurite outgrowth than the linear form. Furthermore, we found that the structure of cyclo-hEF3A is similar to that of the connecting E-F loop region in human laminin alpha3LG4 module by structural analysis using molecular dynamics simulations. These results suggest that the loop structure of the E-F connecting region of the alpha3LG4 module is important for its biological activities. The cyclo-hEF3A peptide may be useful for the development of therapeutic reagents especially for wound healing and nerve regeneration.
authors.affiliation: School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo 192-0392, Japan.
authors.forename: Kozue
authors.initials: K
authors.lastname: Kato-Takagaki
authors.forename: Nobuharu
authors.initials: N
authors.lastname: Suzuki
authors.forename: Fumiharu
authors.initials: F
authors.lastname: Yokoyama
authors.forename: Shu
authors.initials: S
authors.lastname: Takaki
authors.forename: Koji
authors.initials: K
authors.lastname: Umezawa
authors.forename: Junichi
authors.initials: J
authors.lastname: Higo
authors.forename: Mayumi
authors.initials: M
authors.lastname: Mochizuki
authors.forename: Yamato
authors.initials: Y
authors.lastname: Kikkawa
authors.forename: Shinya
authors.initials: S
authors.lastname: Oishi
authors.forename: Atsushi
authors.initials: A
authors.lastname: Utani
authors.forename: Motoyoshi
authors.initials: M
authors.lastname: Nomizu
mesh_terms: D000595:Amino Acid Sequence; D002448:Cell Adhesion; D002460:Cell Line; D006493:Heparin / Q000378:metabolism; D006801:Humans; D007797:Laminin / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016501:Neurites / Q000187:drug effects / Q000502:physiology; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D010456:Peptides, Cyclic / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship; D053671:Syndecan-4 / Q000378:metabolism
chemical_list: D007797:Laminin; D009842:Oligopeptides; D010456:Peptides, Cyclic; D011994:Recombinant Proteins; D053671:Syndecan-4; C518304:cyclo-hEF3A peptide; C095567:laminin alpha 3; D006493:Heparin