Interactions of cations with membrane fractions of smooth and rough strains of Salmonella typhimurium and other Gram-negative bacteria.

Journal of Bacteriology
H Stan-Lotter, K E Sanderson

Abstract

Addition of cations (20 to 50 mM for Mg(2+) or Ca(2+) or 100 to 500 mM for Na(+)) to N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid buffer during preparation of membranes from smooth and rough strains of Salmonella typhimurium LT2, Salmonella minnesota, and Escherichia coli O8 had two effects on the composition of the membranes isolated. First, in rough strains of chemotypes Ra to Re the "total membranes" (pellets from high-speed centrifugation) were deficient in the proteins of the outer membrane. The missing proteins were found to have been sedimented in a prior low-speed centrifugation in a fraction we call "cation-aggregated membranes." Since these membranes were enriched for lipopolysaccharide and for outer membrane proteins, deficient in succinic dehydrogenase, and contained primarily the dense peak after sucrose gradient centrifugation, it appears to be relatively pure outer membrane. About 10% of the membrane protein of smooth strains and up to 50% that of rough strains were cation-aggregated membranes, appearing to contain most of the outer membrane of rough strains. Thus, cation aggregation may be a useful means of preparation of outer membrane samples. The second effect was that with cation addition, several hig...Continue Reading

References

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