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Identification of a heparin binding site and the biological activities of the laminin α1 chain carboxy‐terminal globular domain
Author(s) -
Yoshida Ichiro,
Tashiro KenIchiro,
Monji Akira,
Nagata Isao,
Hayashi Yoshihito,
Mitsuyama Yoshio,
Tashiro Nobutada
Publication year - 1999
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/(sici)1097-4652(199904)179:1<18::aid-jcp3>3.0.co;2-k
Subject(s) - laminin , neurite , cell adhesion , adhesion , peptide , microbiology and biotechnology , neural cell adhesion molecule , chemistry , integrin , biochemistry , binding domain , binding site , biology , cell , in vitro , organic chemistry
The carboxy‐terminal globular domain (G‐domain) of the laminin α1 chain has been shown to promote heparin binding, cell adhesion, and neurite outgrowth. In this study, we defined the potential sequences originating from the G‐domain of laminin α1 chain which possess these functional activities. A series of peptides were synthesized from the G‐domain, termed LG peptides (LG‐1 to LG‐6) and were tested for their various biological activities. In the direct [ 3 H]heparin binding assays, LG‐6 (residues 2,335–2,348: KDFLSIELVRGRVK) mediated high levels of [ 3 H]heparin binding, and this peptide also directly promoted cell adhesion and spreading, including B16F10, M2, HT1080, and PC12 cells. The peptide LG‐6 also promoted the neurite outgrowth of PC12 cells, mouse granule cells, and chick telencephalic cells. An anti‐peptide LG‐6 antibody inhibited laminin‐1 and peptide LG‐6–mediated cell adhesion and neurite outgrowth. Furthermore, an anti‐integrin α2 antibody also inhibited the cell adhesion activity. These results suggest that peptide LG‐6 plays a functional role as a heparin binding site in the G‐domain of the laminin α1 chain, and this sequence was thus concluded to play a crucial role in regulating cell adhesion and spreading and neurite outgrowth which is related to integrin α2. J. Cell. Physiol. 179:18–28, 1999. © 1999 Wiley‐Liss, Inc.