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Subcellular localization of PhaZc determined by using a sucrose density gradient. Sonicates from PHB-rich cells were fractionated by ultracentrifugation in a sucrose gradient (1 to 2 M; total volume, 11 ml). Each fraction (1.1 ml) was collected and analyzed by Western blotting and immunostaining with antiserum against PhaZc, PhaZ1, or 3HB dehydrogenase (3HBDH); 100% corresponded to PHB and protein concentrations of 65 and 3.7 mg/ml, respectively.
