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. Author manuscript; available in PMC: 2010 Feb 10.
Published in final edited form as: J Biol Chem. 2007 Aug 16;282(40):29504. doi: 10.1074/jbc.M705825200

FIGURE 10. Endogenous Myo1e from oocytes and in vitro translated Myo1e bind to recombinant Csp immobilized on amylose resin.

FIGURE 10

A, extracts from 20 oocytes were prepared as in Fig. 9 and incubated 16 h with 0.1 mg of MBP bound to amylose resin or 0.1 mg of MBP-Csp bound to amylose resin. The resins were washed with buffer A and extracted with SDS sample buffer for Myo1e immunoblot analysis. The extract lane reflects the Myo1e immunoreactivity in extract from a single oocyte. B, equivalent amounts (0.1 mg) of MBP or MBP-Csp were bound to amylose resin and incubated in a solution containing 20 μl of in vitro translated (IVT) Myo1e, Myo1e-head, or Myo1c (see “Experimental Procedures”). After washing, the resin was extracted with SDS sample buffer and subjected to immunoblot analysis. The in vitro translated (IVT) lanes show the immunoreactive Myo1e, Myo1e-head (~75 kDa) or Myo1c (~105 kDa), in 2.5 μl of the in vitro translation reaction. None of these proteins bound to amylose resin with MBP. Only Myo1e was detected bound to resin containing MBP-Csp.