Heterogeneity in lipid composition of the outer membrane and cytoplasmic membrane and cytoplasmic membrane of Pseudomonas BAL-31

The outer membranes and cytoplasmic membranes of the marine bacterium Pseudomonas BAL-31 were separated by washing the cells three times in 0.5 M NaCl and twice in 0.5 M sucrose. Electron microscopy during the removal of membranes revealed that the outer membranes fragmented in a regular manner to g...

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Bibliographic Details
Published in:Journal of bacteriology 1974-09, Vol.119 (3), p.1006-1018
Main Authors: Diedrich, D L, Cota-Robles, E H
Format: Article
Language:English
Subjects:
Autoradiography
Cell Membrane - analysis
Cell Wall - analysis
Chromatography, Thin Layer
Culture Media
Drug Resistance, Microbial
Fatty Acids - analysis
Keto Acids - analysis
Lipids - analysis
Microscopy, Electron
Phosphatidylethanolamines - analysis
Phospholipids - analysis
Phosphorus - analysis
Phosphorus Radioisotopes
Pseudomonas - analysis
Pseudomonas - drug effects
Seawater
Sodium Chloride
Sucrose
Surface-Active Agents - pharmacology
Water Microbiology
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Summary:The outer membranes and cytoplasmic membranes of the marine bacterium Pseudomonas BAL-31 were separated by washing the cells three times in 0.5 M NaCl and twice in 0.5 M sucrose. Electron microscopy during the removal of membranes revealed that the outer membranes fragmented in a regular manner to give rise to fairly uniform vesicles measuring approximately 140 nm in diameter. Isolated outer membranes had a buoyant density in sucrose of 1.230 g per cm(3), whereas the cytoplasmic membranes had a density of 1.194 g per cm(3). The removal of the outer membrane during the application of this procedure was monitored by measuring the release of 2-keto-3-deoxyoctulosonic acid and phospholipid. The cells lost 85.5% of their 2-keto-3-deoxyoctulosonic acid and 47.3% of their phospholipid during this treatment. Complete recovery of outer membrane material could be achieved. The removal of 25.5% of the 2-keto-3-deoxyoctulosonic acid and 0.9% of the phospholipid rendered the cells sensitive to lysis with Triton X-100. The phospholipid composition of the outer membrane was calculated to be 78.9% phosphatidylethanolamine and 16.1% phosphatidylglycerol. The phospholipid composition of the cytoplasmic membrane proved to be 71.5% phosphatidylethanolamine and 23.5% phosphatidylglycerol. The fatty acid composition was also found to be quantitatively heterogeneous between the two membranes.
ISSN:0021-9193