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Identification of laminin α 5 short arm peptides active for endothelial cell attachment and tube formation
Author(s) -
Kikkawa Yamato,
Sugawara Yumika,
Harashima Nozomi,
Fujii Shogo,
Ikari Kazuki,
Kumai Jun,
Katagiri Fumihiko,
Hozumi Kentaro,
Nomizu Motoyoshi
Publication year - 2017
Publication title -
journal of peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 66
eISSN - 1099-1387
pISSN - 1075-2617
DOI - 10.1002/psc.2987
Subject(s) - laminin , endothelial stem cell , peptide sequence , cell adhesion , basement membrane , peptide , human umbilical vein endothelial cell , biochemistry , extracellular matrix , amino acid , glycoprotein , biology , cell , microbiology and biotechnology , integrin , chemistry , in vitro , gene
Laminin‐511, a major component of endothelial basement membrane, consists of α 5, β 1, and γ 1 chains. The short arm region of the α 5 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 α 5 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 α 5LN‐LEa domain. Further, we synthesized 70 peptides covering the amino acid sequences of the α 5LN‐LEa domain. Of the 70 peptides, A5‐16 (mouse laminin α 5 230–243: 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.