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Comparison of the Biochemistry and Rates of Synthesis of Mesosomal and Peripheral Membranes in Bacillus subtilis¹

^a Department of Biology, Georgetown University, Washington, D.C. 20007

ABSTRACT

The membrane vesicle (beaded chain) portion of the mesosomes and peripheral (ghost) membrane of Bacillus subtilis were obtained by protoplast lysis and separated by differential and sucrose gradient centrifugation. Electron microscopy revealed that both fractions were satisfactorily homogeneous. Comparison of the two membrane preparations showed that they were similar with respect to total protein, total phosphorus, and lipid-soluble phosphorus content. Their protein patterns on acrylamide gel electrophreograms did not differ significantly. A possible point of distinction was revealed by a difference spectrum analysis of their cytochromes. The two preparations showed clear quantitative differences in all five of the enzyme activities assayed. Acrylamide gel electrophreograms of peripheral membrane stained for malate dehydrogenase showed four weak isozyme bands, whereas electrophreograms of mesosome membranes exhibited a single strong peak. (A survey of published data on enzymes in mesosome fractions shows a marked lack of correspondence between different species of bacteria.) Comparison of ³H-acetate incorporation into the two membrane fractions showed that both were labeled at the same rate. Similarly, ³⁵SO₄ was taken up by both fractions at a comparable rate and was chased from both comparably. Lipid and protein labeling thus indicates that mesosome vesicle membrane is not a precursor or special growing point of peripheral membrane.

² Present address: Laboratory of Biology of Viruses, NIAID, National Institutes of Health, Bethesda, Md. 20014.

¹ This work was carried out by C.T.P. in partial fulfillment of the requirements of Georgetown University for the degree of Doctor of Philosophy.