This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Johnston, K. H. Articles by Gotschlich, E. C. Search for Related Content PubMed PubMed Citation Articles by Johnston, K. H. Articles by Gotschlich, E. C.

Isolation and Characterization of the Outer Membrane of Neisseria gonorrhoeae

¹ The Rockefeller University, New York, New York 10021

ABSTRACT

The cell envelope of Neisseria gonorrhoeae strain 2686, colonial type 4, was isolated from spheroplasts formed by the action of ethylenediaminetetraacetic acid and lysozyme. Isopycnic centrifugation of osmotically ruptured spheroplasts resolved the cell envelope into two main membrane fractions. Chemical and enzymatic analyses were used to characterize these isolated membranes. Succinic dehydrogenase, reduced nicotinamide adenine dinucleotide oxidase, and D-lactate dehydrogenase were localized in the membrane fraction of buoyant density, ° = 1.141 g/cm³. Lipopolysaccharide and over half of the cell envelope protein were associated with the membrane that banded in sucrose at ° = 1.219 g/cm³. These fractions were consequently designated cytoplasmic and outer or L-membrane, respectively. Sodium dodecyl sulfate-polyacrylamide electrophoresis of isolated membranes demonstrated the relative simplicity of the protein spectrum of the outer membrane. The majority of the protein in this membrane could be accounted for by proteins of molecular weights 34,500, 22,000, and 11,500. The protein of molecular weight 34,500 accounted for 66% of the total protein of the L-membrane. Isoelectric precipitation at pH 4.6 with 10% acetic acid selectively removed this protein from a 150 mM NaCl in 10 mM tris(hydroxymethyl)aminomethane-hydrochloride, pH 7.4, extract of purified outer membrane. At pH 4.0, the other proteins of the L-membrane were precipitated. It was concluded that the membrane components of the cell envelope of N. gonorrhoeae were similar to those of other gram-negative bacteria. The cell envelope fractions described here, in particular the outer membrane, are sufficiently well defined to provide a valuable tool for future biochemical and immunological studies on N. gonorrhoeae.

This article has been cited by other articles:

• Bagchi, A K, Sinha, A K (2005). Phosphotidylinositol-3 kinase-mediated signals in mice immunized with the 57 kDa major antigenic outer-membrane protein of Shigella dysenteriae type 1. J Med Microbiol 54: 631-637 [Abstract] [Full Text]  
• Singleton, T. E., Massari, P., Wetzler, L. M. (2005). Neisserial Porin-Induced Dendritic Cell Activation Is MyD88 and TLR2 Dependent. J. Immunol. 174: 3545-3550 [Abstract] [Full Text]  
• Sinha, A K, Bagchi, A K (2004). Role of anti-CD3 in modulation of Th1-type immune response in Shigella dysenteriae infection. J Med Microbiol 53: 1075-1081 [Abstract] [Full Text]