Figure 8.

MONaKA is associated with the plasma membrane. A, Brain cytoplasmic and membrane fractions were analyzed by Western blot to determine the distribution of MONaKA (MO). MONaKA protein is found in both the brain cytoplasm and membrane fractions, with approximately two-thirds of the protein associated with the membrane based on densitometric analysis. B, Cell surface biotinylation. tsA201 cells were transfected with Myc-Na,K-ATPase β1 subunit (lane 1) or HA-MONaKA (lane 2) or both together (lane 3). After surface biotinylation (see Materials and Methods), both MONaKA and the β1 subunit (positive control) can be detected in avid in precipitates (bottom panels) using anti-HA or -Myc antibodies, respectively. The cytoplasmic protein Myc-14-3-3 is not present in the avidin precipitate under these experimental conditions (lane 4).