FIGURE 3.

mAKAPβ binds AC5 directly. A, AC5 has 12 transmembrane spans (hatched bars) and an N-terminal (5NT) and two catalytic (5C1 and 5C2) intracellular domains. B, cell lysates (500 μg) from COS-7 cells transfected with a mAKAPβ or control expression vector were incubated with 30 μg of GST or GST-5NT (1 μm) in the presence or absence of His₆-tagged 5C1 and 5C2 (2 μm each) and 100 μm fsk. Complexes were precipitated with glutathione resin. Western blots for mAKAPβ and inputs are shown. 50% of the starting material is shown for mAKAPβ input. n = 3. C, His₆-tagged 5C1 and 5C2 (1 μm) were incubated with 100 μm fsk and GST or GST-mAKAP fusion proteins and precipitated with glutathione resin. n = 3.