In contrast to the ^-786 T-variant, the ^-786 C-variant of the endothelial nitric oxide synthase (~nos-3) gene is shear stress-insensitive and associated with coronary artery disease. We here compare changes in shear stress-induced protein expression in human endothelial cells (HUVEC) with ~nos-3 ^-786 T/T and -786 C/C genotype. Primary genotyped HUVEC were subjected to shear stress (24 h; 30 dyne/cm²). Total protein was separated by standard 2D-gel electrophoresis and protein expression was compared to control cells by densitometry. Differentially expressed proteins were identified by mass spectrometry. In HUVEC with ^-786T /T genotype 8 out of 28 differentially regulated proteins were structural proteins. In cells with ^-786 C/C genotype, 9 out of 19 differentially expressed proteins were specific for these cells, among those Mn- superoxide dismutase and peroxiredoxin 1. NO bioavailability is one major determinant of endothelial phenotype. Endothelial cells with ~nos-3 ^-786 C/C genotype show a specific response to laminar shear stress aimed at controlling oxidative stress. This might be the first sign of endothelial dysfunction finally resulting in coronary artery disease.