Separation and Characterization of the Outer Membrane of Pseudomonas aeruginosa
- 1 July 1978
- journal article
- research article
- Published by Oxford University Press (OUP) in The Journal of Biochemistry
- Vol. 84 (1), 179-191
- https://doi.org/10.1093/oxfordjournals.jbchem.a132106
Abstract
A method is described for the preparation of outer and cytoplasmic membranes of Pseudomonas aeruginosa, and the outer membrane proteins characterized. Isolated outer and cytoplasmic membranes differed markedly in the content of 2-keto-3-deoxyoctonate (lipopolysaccharide) and phospholipid as well as in the localization of certain enzymes (NADH oxidase, succinate dehydrogenase, D-lactate dehydrogenase, malate dehydrogenase, and phospholipase), and also in the microscopic morphology. The outer membrane preparation showed activity neutralizing a certain bacteriocin or bacteriophages, whereas the cytoplasmic membrane preparation showed no neutralizing activity. The protein composition of membrane preparations from five different strains of P. aeruginosa {P14, M92 (PAO1), PAC1, P15, and M2008 (PAT)} were determined by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. More than 50 protein bands were detected in the cytoplasmic membrane preparation. The protein compositions of outer membranes from the five different strains were very similar: at least 6 major bands were found (apparent molecular weights: Band D, 50,000; band E, 45,000; band F, 33,000; bands G and H, 21,000; and band I, 8,000). The protein composition of outer membranes was affected by some physiological growth conditions. Some features of major outer membrane proteins were also studied. Band F showed anomalous migration on SDS polyacrylamide gel electrophoresis depending on the solubilizing conditions or pretreatment with TCA. Band I seemed to be a protein analogous to the lipoprotein which had been found in the outer membrane of Escherichia coli.Keywords
This publication has 17 references indexed in Scilit:
- Separation of the cytoplasmic and outer membrane of pseudomonas aeruginosa PAO.1Biochemical and Biophysical Research Communications, 1977
- Effects of Heating in Dodecyl Sulfate Solution on the Conformation and Electrophoretic Mobility of Isolated Major Outer Membrane Proteins from Escherichia coli K-121The Journal of Biochemistry, 1976
- Maltose Transport in Escherichia coli K12. A Comparison of Transport Kinetics in Wild-Type and lamba-Resistant Mutants with the Dissociation Constants of the Maltose-Binding Protein as Measured by Fluorescence QuenchingEuropean Journal of Biochemistry, 1976
- Suppression of Fluorophenylalanine Resistance by Mutation to Streptomycin Resistance in Pseudomonas aeruginosaJournal of Bacteriology, 1966
- Acrylamide-Gel Electrophorograms by Mechanical Fractionation: Radioactive Adenovirus ProteinsScience, 1966
- Studies of a PyocinThe Journal of Biochemistry, 1964
- Studies of a PyocinThe Journal of Biochemistry, 1964
- Phosphorus Assay in Column ChromatographyJournal of Biological Chemistry, 1959
- Kinetic studies of pigment synthesis by non‐sulfur purple bacteriaJournal of Cellular and Comparative Physiology, 1957
- PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENTJournal of Biological Chemistry, 1951