title: Identification of laminin α5 short arm peptides active for endothelial cell attachment and tube formation.
abstract: Laminin-511, a major component of endothelial basement membrane, consists of α5, β1, and γ1 chains. The short arm region of the α5 chain is a structural feature of endothelial laminins. In this study, we identified active sequences for human umbilical vein endothelial cells (HUVECs) using recombinant proteins and synthetic peptides. The short arm of the α5 chain contains three globular domains [laminin N-terminal globular domain, laminin 4 domain a, and laminin 4 domain b (LN, L4a, and L4b)] and three rod-like elements [laminin epidermal growth factor-like domain a, b, and c (LEa, LEb, and LEc)]. The cell attachment assay using recombinant proteins showed that RGD-independent cell attachment sites were localized in the α5LN-LEa domain. Further, we synthesized 70 peptides covering the amino acid sequences of the α5LN-LEa domain. Of the 70 peptides, A5-16 (mouse laminin α5 230-243: LENGEIVVSLVNGR) potently exhibited endothelial cell attachment activity. An active sequence analysis using N-terminally and C-terminally truncated A5-16 peptides showed that the nine-amino acid sequence IVVSLVNGR was critical for the endothelial cell attachment activity. Cell adhesion to the peptides was dependent on both cations and heparan sulfate. Further, the A5-16 peptide inhibited the capillary-like tube formation of HUVECs with the cells forming small clumps with short tubes. The eight-amino acid sequence EIVVSLVN in the A5-16 peptide was critical to inhibit HUVEC tube formation. This amino acid sequence could be useful for grafts and thus modulate endothelial cell behavior for vascular surgery. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.
authors.affiliation: Department of Clinical Biochemistry, Tokyo University of Pharmacy and Life Sciences, Tokyo, 192-0392, Japan.
authors.forename: Yamato
authors.initials: Y
authors.lastname: Kikkawa
authors.affiliation: Department of Clinical Biochemistry, Tokyo University of Pharmacy and Life Sciences, Tokyo, 192-0392, Japan.
authors.forename: Yumika
authors.initials: Y
authors.lastname: Sugawara
authors.affiliation: Department of Clinical Biochemistry, Tokyo University of Pharmacy and Life Sciences, Tokyo, 192-0392, Japan.
authors.forename: Nozomi
authors.initials: N
authors.lastname: Harashima
authors.affiliation: Department of Clinical Biochemistry, Tokyo University of Pharmacy and Life Sciences, Tokyo, 192-0392, Japan.
authors.forename: Shogo
authors.initials: S
authors.lastname: Fujii
authors.affiliation: Department of Clinical Biochemistry, Tokyo University of Pharmacy and Life Sciences, Tokyo, 192-0392, Japan.
authors.forename: Kazuki
authors.initials: K
authors.lastname: Ikari
authors.affiliation: Department of Clinical Biochemistry, Tokyo University of Pharmacy and Life Sciences, Tokyo, 192-0392, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Kumai
authors.affiliation: Department of Clinical Biochemistry, Tokyo University of Pharmacy and Life Sciences, Tokyo, 192-0392, Japan.
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Katagiri
authors.affiliation: Department of Clinical Biochemistry, Tokyo University of Pharmacy and Life Sciences, Tokyo, 192-0392, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Hozumi
authors.affiliation: Department of Clinical Biochemistry, Tokyo University of Pharmacy and Life Sciences, Tokyo, 192-0392, Japan.
authors.forename: Motoyoshi
authors.initials: M
authors.lastname: Nomizu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D042783:Endothelial Cells; D057809:HEK293 Cells; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D007797:Laminin / Q000737:chemistry*; D051379:Mice; D010455:Peptides / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D007797:Laminin; D010455:Peptides; D011994:Recombinant Proteins; C097043:laminin alpha5
keywords: cell adhesion; endothelial cell; laminin