Figure 1.

Purification of recombinant wild-type and Ser-329 mutants of Acanthamoeba myosin IC. SDS/PAGE of the total cell extract (200,000 × g supernatant) of cells expressing wild-type AMIC that was applied to the affinity column (Ext) and the purified wild-type AMIC (wt) and mutants S329A (Ala), S329D (Asp), S329E (Glu), and S329N (Asn). The positions of the heavy chain (HC) and light chain (LC) are indicated. Approximately 3 μg of wild-type myosin and 6 μg of the mutant myosins were analyzed. The gels were stained with Coomassie blue. The extract and wild-type myosin were on one gel and the mutant myosins on another. For details, see Materials and Methods.