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. 2009 Jun 3;284(30):20070–20078. doi: 10.1074/jbc.M109.016865

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© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 1. — Purification and kinetics of recombinant SHIP. A, left panel, 100 ng of SHIP1 purified from HEK-293 cells, resolved on an 8% SDS gel and stained with Coomassie Blue. The right panel presents an immunoblot with a SHIP1 antibody. B, upper panel, time-dependent phosphatase activity of SHIP1 with the indicated concentrations of PtdIns 3,4,5-P₃ (PIP₃); lower panel, plot of initial rate against PtdIns 3,4,5-P₃ concentration. V_max and K_m were estimated as 1.8 pmol P_i/min/ng and 46.0 μm, respectively. C, upper panel, time-dependent phosphatase activity of SHIP1 with different concentrations of Ins-1,3,4,5-P₄ (IP₄) as the substrate. V_max and K_m were estimated as 3.8 pmol P_i/min/ng and 351.4 μm, respectively.