The 67-kDa laminin-binding protein is involved in shear stress-dependent endothelial nitric-oxide synthase expression.
It has been suggested that the mechanical forces acting on endothelial cells may be sensed in part by cell-matrix connections. We therefore studied the role of different matrix proteins, in particular laminin I, on a shear stress-dependent endothelial response, namely nitric-oxide synthase (eNOS) expression. Primary porcine aortic endothelial cells were seeded onto glass plates either noncoated (NC cells) or precoated with fibronectin (FN cells), laminin (LN cells), or collagen I (CN cells). Western blots were used to detect differences in the final matrix composition of these cells. A shear stress of 16 dyn/cm2 was applied for 6 h. Only LN cells showed detectable amounts of laminin I in their underlying matrix when they reached confluence. They reacted with a 2-fold increase of eNOS expression (n = 16, p < 0.001) to the exposure of shear stress, which went along with enhanced eNOS protein and NO release. In contrast, neither FN cells (n = 9) nor NC cells (n = 13) showed a significant increase of eNOS expression under shear stress. The increase in CN cells was borderline (1.4-fold; n = 9, p < 0.05) and was not associated with an increase of eNOS protein. The shear-induced increase in eNOS expression of LN cells was abolished by the peptide YIGSR, which blocks the cellular binding to laminin I via a 67-kDa laminin-binding protein, whereas a control peptide (YIGSK) had no effect. The induction of eNOS expression by shear stress is stimulated by an interaction of endothelial cells with laminin which is, at least in part, mediated by a 67-kDa laminin-binding protein.
Gloe
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Institute of Physiology, Ludwig Maximilians University, Schillerstrasse 44, D 80336 Munich, Germany. Gloe@lrz.uni-muenchen.de
Riedmayr
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S
Sohn
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HY
Pohl
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U
D000818:Animals; D015153:Blotting, Western; D002448:Cell Adhesion; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D004730:Endothelium, Vascular / Q000201:enzymology*; D005353:Fibronectins / Q000378:metabolism; D007797:Laminin / Q000378:metabolism; D008970:Molecular Weight; D019001:Nitric Oxide Synthase / Q000096:biosynthesis*; D052250:Nitric Oxide Synthase Type III; D009842:Oligopeptides / Q000494:pharmacology; D011498:Protein Precursors; D017474:Receptors, Laminin / Q000502:physiology*; D013314:Stress, Mechanical; D013552:Swine
D005353:Fibronectins; D007797:Laminin; D009842:Oligopeptides; D011498:Protein Precursors; D017474:Receptors, Laminin; C081571:tyrosyl-isoleucyl-glycyl-seryl-arginine; D003094:Collagen; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III