The outer membrane of Campylobacter coli, C. jejuni and C. fetus cell envelopes appeared as three fractions after sucrose gradient centrifugation. Each outer membrane fraction was contaminated with succinate dehydrogenase activity from the cytoplasmic membrane fraction. Similarly the inner membrane fraction was contaminated with 2-ketodeoxyoctonate and outer membrane proteins including the porin(s). The separation of these two membranes was not facilitated by variations in lysozyme treatment, cell age, presence or absence of flagella, or longer lipopolysaccharide chain length. Sodium lauroyl sarcosinate extraction resulted in an outer membrane fraction which contained some inner membrane contamination and produced multiple bands upon sucrose gradient centrifugation. Triton X-100 extraction removed the inner membrane from the outer membrane and Triton X-100/EDTA treatment extracted lipopolysaccharide-rich regions of the outer membrane which contained almost exclusively the Campylobacter porin(s). These data indicated that the inner and outer membranes of the Campylobacter cell envelope were very difficult to separate, possibly because of extensive fusions between these two membranes.
AmesG.F.-L.,
SpudichE. N.,
NikaidoH.1974; Protein composition of the outer membrane of Salmonella typhimurium: effect of lipopolysaccharide mutations. Journal of Bacteriology 117:406–416
BayerM. E.1979; The fusion sites between outer membrane and cytoplasmic membrane of bacteria: their role in membrane assembly and virus infection. In Bacterial Outer Membranes. Biogenesis and Functions pp. 167–202InouyeM.
Edited by New York: John Wiley;
BradfordM. M.1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Chemistry 72:248–254
BuckG. E.,
ParshallK. A.,
DavisC. P.1983; Electron microscopy of the coccoid form of Campylobacter jejuni
. Journal of Clinical Microbiology 18:420–421
FairbanksG.,
SteckT. L.,
WallachD. F. H.1971; Electrophoretic analysis of major polypeptides of the human erythrocyte membrane. Biochemistry 10:2606–2616
FilipC.,
FletcherG.,
WulffJ. L.,
EarhartC. F.1973; Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate. Journal of Bacteriology 115:717–722
HancockR. E. W.,
NikaidoH.1978; Outer membranes of Gram-negative bacteria. XIX. Isolation from Pseudomonas aeruginosaPAOl and use in reconstitution and definition of the permeability barrier. Journal of Bacteriology 136:381–390
KoplowJ.,
GoldfineH.1974; Alterations in the outer membrane of the cell envelope of heptose- deficient mutants of Escherichia coli
. Journal of Bacteriology 117:527–543
LoganS. M.,
TrustT. J.1984; Structural and antigenic heterogeneity of lipopolysaccharides of Campylobacter jejuni and Campylobacter coli
. Infection and Immunity 45:210–216
MoranA. P.,
UptonM. E.1986; A comparative study of the rod and coccoid forms of Campylobacter jejuni ATCC 29428. Journal of Applied Bacteriology 60:103–110
MoranA. P.,
UptonM. E.1987; Factors affecting production of coccoid forms of Campylobacter jejuni on solid media during incubation. Journal of Applied Bacteriology 62:527–537
NewellD. G.,
McbrideH.,
PearsonA. D.1984; The identification of outer membrane proteins and flagella of Campylobacter jejuni
. Journal of General Microbiology 130:1201–1208
NgL.-K.,
SherburneR.,
TaylorD. E.,
StilesM. E.1985; Morphological forms and viability of Campylobacter species studied by electron microscopy. Journal of Bacteriology 164:338–343
OsbornM. J.,
GanderJ. E.,
ParisiE.,
CarsonJ.1972; Mechanism of assembly of the outer membrane of Salmonella typhimurium: isolation and characterisation of cytoplasmic and outer membrane. Journal of Biological Chemistry 247:3962–3972
PageW. J.,
Von TigerstromM.1982; Iron- and molybdenum-repressible outer membrane proteins in competent Azotobacter vinelandii
. Journal of Bacteriology 151:237–242
PerezG. I. P.,
HopkinsJ. A.,
BlaserM.1985; Antigenic heterogeneity of lipopolysaccharides from Campylobacter jejuni and Campylobacter fetus
. Infection and Immunity 48:528–533
RollinsD. M.,
ColwellR. R.1986; Viable but non culturable stage of Campylobacter jejuni and its role in survival in the natural aquatic environment. Applied and Environmental Microbiology 52:531–538
SchnaitmanC. A.1971; Effect of ethylenediamine- tetraacetic acid, Triton X-100, and lysozyme on the morphology and chemical composition of isolated cell walls of Escherichia coli
. Journal of Bacteriology 108: 553–563
SmitJ.,
NikaidoH.1978; Outer membrane of Gram-negative bacteria. XVIII. Electron microscopic studies on porin insertion sites and growth of cell surface of Salmonella typhimurium
. Journal of Bacteriology 135:687–702
TaylorD. E.,
GarnerR. S.,
AllanB. J.1983; Characterization of tetracycline resistance plasmids from Campylobacter jejuni and Campylobacter coli
. Antimicrobial Agents and Chemotherapy 24:930–935
WhiteD. A.,
LennarzW. J.,
SchnaitmanC. A.1972; Distribution of lipids in the wall and cytoplasmic membrane subfractions of the cell envelope of Escherichia coli
. Journal of Bacteriology 109: 686–690