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. 2011 Aug 16;286(39):33729–33736. doi: 10.1074/jbc.M111.291591

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© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 1. — Purification and properties of ACSF3. A, Western analysis of bovine and mouse mitochondrial extracts probed with antibody against ACSF3. Lanes 1 and 3, 60 μg of mitochondrial protein from bovine and mouse heart extracts, respectively; lanes 2 and 4, 10 μg of protein from partially purified bovine and mouse preparations, respectively. B, general protein staining of partially purified bovine (lane 1) and mouse (lane 2) ACSF3 preparations. C, General protein staining of recombinant ACSF3 protein preparations. Lane 1, purified GST-ACSF3; lane 2, GST-ACSF3 after treatment with tobacco etch virus protease; lanes 3–5, final purified preparations of wild-type ACSF3 and mutants R354A and R354L, respectively. D, malonyl-CoA synthetase activity of the ACSF3 preparation. Assays were performed at 37 °C for 30 min, and reaction components were separated by HPLC.