Figure 5. SDS/PAGE analysis of GAHp–His₆, Gfa1p and Gfa1p^S208A phosphorylation by PKA.

Mixtures obtained after a 30 min incubation of a given protein with indicated components were separated by SDS/PAGE. (A) Gel stained with the GelCode™ kit. (B) The same gel as (A) stained with Coomassie Brilliant Blue. Lane 1, GAHp–His₆ and PKA; lane 2, GAHp–His₆; lane 3, GAHp–His₆, PKA and H-89; lane 4, Gfa1p and PKA; lane 5, Gfa1p, PKA and 10 mM Fru-6-P; lane 6, S208AGfa1p and PKA; lane 7, Gfa1p. Each mixture contained 10 μM cAMP, 1 mM ATP, 10 mM EDTA and 40 mM NaF. Protein load: 10 μg (lanes 1–3) and 3 μg (lanes 4–7). M, molecular mass markers (from top to bottom): 120 kDa, 100 kDa, 85 kDa, 70 kDa, 60 kDa, 50 kDa and 40 kDa.