Laminin-10 and Lutheran blood group glycoproteins in adhesion of human endothelial cells.
Laminin alpha5-chain, a constituent of laminins-10 and -11, is expressed in endothelial basement membranes. In this study we evaluated the roles of alpha5 laminins and Lutheran blood group glycoproteins (Lu), recently identified receptors of the laminin alpha5-chain, in the adhesion of human dermal microvascular and pulmonary artery endothelial cells. Field emission scanning electron microscopy and immunohistochemistry showed that the endothelial cells spread on laminin-10 and formed fibronectin-positive fibrillar adhesion structures. Immunoprecipitation results suggested that the cells produced fibronectin, which they could use as adhesion substratum, during the adhesion process. When the protein synthesis during the adhesion was inhibited with cycloheximide, the formation of fibrillar adhesions on laminin-10 was abolished, suggesting that laminin-10 does not stimulate the formation of any adhesion structures. Northern and Western blot analyses showed that the cells expressed M(r) 78,000 and 85,000 isoforms of Lu. Quantitative cell adhesion assays showed that in the endothelial cell adhesion to laminin-10, Lu acted in concert with integrins beta(1) and alpha(v)beta(3), whereas in the adhesion to laminin-10/11, Lu and integrin beta(1) were involved. In the cells adhering to the alpha5 laminins, Lu and the integrins showed uniform cell surface distribution. These findings indicate that alpha5 laminins stimulate endothelial cell adhesion but not the formation of fibrillar or focal adhesions. Lu mediates the adhesion of human endothelial cells to alpha5 laminins in collaboration with integrins beta(1) and alpha(v)beta(3).
Vainionpää
Noora
N
Institute of Biomedicine/Anatomy, PO Box 63, Haartmaninkatu 8, University of Helsinki, FIN-00014 Helsinki, Finland. noora.vainionpaa@helsinki.fi
Kikkawa
Yamato
Y
Lounatmaa
Kari
K
Miner
Jeffrey H
JH
Rousselle
Patricia
P
Virtanen
Ismo
I
D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D016023:Integrins / Q000378:metabolism; D007797:Laminin / Q000235:genetics / Q000378:metabolism*; D008188:Lutheran Blood-Group System; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D016608:Talin / Q000378:metabolism; D016596:Vinculin / Q000378:metabolism
C090228:BCAM protein, human; D015815:Cell Adhesion Molecules; D005353:Fibronectins; D016023:Integrins; D007797:Laminin; D008188:Lutheran Blood-Group System; D009363:Neoplasm Proteins; D016608:Talin; C119013:laminin 10; D016596:Vinculin