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. 1997 Nov 25;94(24):13317–13322. doi: 10.1073/pnas.94.24.13317

Figure 2.

Figure 2

Immunolocalization and biochemical detection of a Gi protein with ZGs from rat exocrine pancreas. (a and b) Immunoblot analysis. Identification of Gαi3-immunoreactive antigen associated with ZGM. (a) Ten micrograms each of TH and ZGM was resolved using a single dimension 12.5% SDS gel electrophoresis before electrotransfer on to nitrocellulose and immunoblotting. A 44-kDa immunoreactive band was detected in both fractions but enriched in the ZGM. (b) Twenty five micrograms of ZGM protein was resolved on two-dimensional 16-BAC gel electrophoresis before electrotransfer and immunoblotting using the Gαi3-specific antibody. Note a single spot at ≈44 kDa, suggesting that only one Gαi3 isoform is present in the ZGM. (c) Detection of Gi-specific GTPase activity associated with ZG. GTPase activity in 25 μg of ZG incubated at 30°C for 5 min in the presence of varying concentrations of either Mas7 or Mas17 demonstrates a dose-dependent increase in activity in the presence of Mas7, which is absent in the presence of Mas17. Values are mean ± SE.