Membranes obtained from whole-cell lysates of Burkholderia (Pseudomonas) pseudomallei (strain 319a) were separated into four fractions by sucrose density gradient centrifugation. Membranes were characterized by enzymic and chemical analyses, and by SDS-PAGE. Cytoplasmic membranes and two forms of outer membranes (OM-1, OM-2) were detected. The major outer-membrane proteins had M r values of 70000, 38000, 31000, 24000 and 17000. To determine which outer-membrane proteins were common to B. pseudomallei strains, OM-1 fractions from 12 different strains were prepared. SDS-PAGE analysis of these fractions demonstrated that the five major outer-membrane proteins were common to the strains tested. Further studies have shown that an M r 110000 protein, which is oligomeric in that it migrates as an M r 38000 protein upon heating at 95 °C and which is peptidoglycan-associated, serves as a porin in B. pseudomallei. Using proteoliposomes reconstituted from this protein and phospholipid, it was demonstrated by the liposome-swelling assay that this protein acts as a porin through which small saccharides may diffuse. Further characterization of this M r 38000 protein will be important in delineating the role of this molecule in the permeability of the B. pseudomallei outer membrane.
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