Separation and characterization of membranes from vegetative, Myxoccocus xanthus DK1622 cells
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The inability to separate inner and outer membranes in wild-type Myxoccocus xanthus DK1622 has been a major obstacle in examining the physiology of this organism. Using spheroplasting in combination with sucrose density centrifugation we now report isolation and characterization of several membrane fractions. Biochemical analysis suggests that these fractions correspond to inner, hybrid and outer membranes. A fifth membrane fraction contains a Ta-1 polyketide synthetase. Two dimensional SDS-PAGE gel analysis indicated the presence of distinct inner and outer membrane proteomes. Using this approach in conjunction with Western blotting, we demonstrate differential localization of several surface-associated proteins required for social motility and signaling.