Fig. 2.

Purification of PcaK. (a) Cell fractions and column purification fractions were loaded onto SDS–PAGE gels and stained with Coomassie. (b) Equivalent gel subject to Western blotting with anti-V5. Purified PcaK migrated as two bands corresponding to the apparent molecular weights of PcaK monomer (32 kDa) and dimer (70 kDa). Fractions are: Whole cell, total cell lysate; Cytoplasm, supernatant after ultracentrifugation; Membranes, pellet after ultracentrifugation; DDM-soluble, membranes solubilized in detergent DDM, supernatant after ultracentrifugation; Unbound, IMAC column flow-through; Wash, column wash fraction; Elution, column eluent. M, molecular weight marker in kDa as shown. μg protein, μg total protein loaded per lane. BF, buffer front.