Plasmids transfected . | Basal activity . | Activity with 125 μmol l-1 SNP . | Stimulation . |
---|---|---|---|
10 μg MsGC-α1+10 μg MsGC-β1 | 7.72±0.72 | 43.10±0.96 | 5.6-fold |
10 μg MsGC-α1+10 μg MsGC-β1+10 μg MsGC-β3 | 4.63±0.73 | 16.10±1.69 | 3.5-fold |
10 μg MsGC-α1+10 μg MsGC-β1+20 μg MsGC-β3 | 7.72±0.24 | 13.80±1.27 | 1.8-fold |
10 μg MsGC-β3 | 11.10±1.00 | 9.63±0.43 | 0.9-fold |
20 μg MsGC-β3 | 24.4±0.10 | 22.1±1.10 | 0.9-fold |
Plasmids transfected . | Basal activity . | Activity with 125 μmol l-1 SNP . | Stimulation . |
---|---|---|---|
10 μg MsGC-α1+10 μg MsGC-β1 | 7.72±0.72 | 43.10±0.96 | 5.6-fold |
10 μg MsGC-α1+10 μg MsGC-β1+10 μg MsGC-β3 | 4.63±0.73 | 16.10±1.69 | 3.5-fold |
10 μg MsGC-α1+10 μg MsGC-β1+20 μg MsGC-β3 | 7.72±0.24 | 13.80±1.27 | 1.8-fold |
10 μg MsGC-β3 | 11.10±1.00 | 9.63±0.43 | 0.9-fold |
20 μg MsGC-β3 | 24.4±0.10 | 22.1±1.10 | 0.9-fold |
COS-7 cells were transfected with the plasmids as shown plus a plasmid coding for β-galactosidase and assayed for guanylyl cyclase activity in the presence or absence of 125 μmol l-1 sodium nitroprusside(SNP) as described in Materials and methods, except that the MnCl2was replaced by 4 mmol l-1 MgCl2.
Extracts were also assayed for β-galactosidase activity and the guanylyl cyclase activity shown for each plasmid combination was normalized to the β-galactosidase activity present in the vector-transfected sample.
Activity levels are expressed as pmoles cGMP formed min-1mg-1 protein.
Values are means ± S.E.M. of 3 determinations.