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. 2012 Aug 22;287(41):34635–34645. doi: 10.1074/jbc.M112.357509

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© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 1. — PGAM5 is an IMM-resident protein phosphatase. A and B, biochemical characterization of PGAM5 in mitochondria is shown. Isolated mitochondria from HEK293A cells treated with the indicated concentrations of trypsin in the indicated buffer conditions were solubilized and subjected to immunoblotting (IB) (A). The isolated membrane fraction from mouse liver mitochondria was subjected to sucrose density gradient centrifugation followed by IB (B). C, ultrastructural characterization of the localization of PGAM5 in mitochondria is shown. HeLa-PGAM5-FLAG cells were subjected to ordinary electron microscopy (left panel) and immunoelectron microscopy using ultrathin cryosections (right panel). Most of the 10-nm gold particles representing PGAM5-FLAG are mostly localized along mitochondrial cristae. Scale bars: 1 μm (left panel) and 0.5 μm (right panel). D, shown is a schematic view of the localization of the mitochondrial proteins examined in A and B. PGAM5 is mainly localized in the IMM with the phosphatase domain facing the intermembrane space.