Table 1.

cAMP-elevating (iloprost) and cGMP-elevating (riociguat, various NO donors) platelet inhibitors regulate VASP and ENSA phosphorylation.

Gene	Uniprot	P-Site	Av. Ratio Iloprost	Av. Ratio DEA-NO	Av. Ratio Riociguat	Av. Ratio SNC	Av. Ratio SNP	Copy Number/Platelet
VASP	P50552	S239	7.17	12.19	9.22	7.67	6.93	44,600
ENSA	O43768	S109	17.97	14.04	16.03	13.88	12.46	7800
ENSA	O43768	S67	0.53	0.82	0.62	0.72	0.82	7800

Washed human platelets were incubated without (control) or with the following test substances : iloprost (5 nM, 2 min); diethylamine NONOate (DEA-NO 5 µM, 2 min.); riociguat (10 µM, 5 min); sodium S-nitrosocysteine (SNC, 5 µM, 2 min) or sodium nitroprusside (SNP, 5 µM, 2 min). The reaction was stopped with 4× lysis buffer, samples were shock frozen in liquid nitrogen and analysed by quantitative phosphoproteomics as described [40]. The change of various phosphosites in response to the test substance compared to control (increase or decrease) is shown as average ratio (av. ratio). The copy number indicates the number of molecules of a given protein per platelet obtained in earlier studies [34].