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. 2009 Oct 6;284(48):33571–33579. doi: 10.1074/jbc.M109.055897

FIGURE 4.

FIGURE 4.

Interaction of Aurora kinases with Par3. A, schematic representations of Aurora A and B, Par3, and individual domains of Par3 (CR, CR1, conserved region 1, amino acids 1–200; P1–3, PDZ domains 1–3, P1, amino acids 200–400, P2, 400–560; P3, 560–780; PB, aPKCBD, 700–1337) are shown. The numbers refer to the positions in the amino acid sequence. B, bacterially expressed MBP fusion proteins of different Par3 domains were coupled to amylose beads as indicated and incubated with lysates of HEK 293T cells transfected with the expression for GFP-tagged Aurora A or Aurora B (Inp, input). The expression of comparable amounts of protein was confirmed by Western blotting (WB) using anti-MBP and anti-GFP antibodies. Numbers indicate molecular mass in kDa. C, bacterially expressed GST fusion proteins of different Par3 domains (CR, P1, P2, P3, and PB) were coupled to glutathione-Sepharose beads as indicated and incubated with bacterially expressed MBP fusion proteins of the N-terminal (Aurora A–N, amino acids 1–133) or kinase domains of Aurora A (Aurora A–K, amino acids 133–403) or B (Aurora B–K, amino acids 83–344). The expression of comparable amounts of protein was confirmed by Western blotting using anti-MBP and anti-GST antibodies. Numbers indicate molecular mass in kDa.