Cytochrome C Reductase Activity of Meningopneumonitis Organisms at Different Stages of Development

Tamura, Akira; Manire, G. P.

doi:10.3181/00379727-129-33328

Cited by 7 publications

(4 citation statements)

References 2 publications

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“…To isolate and purify MP39.5, a common approach would be to first obtain pure chlamydial outer membranes and then extract the protein from these preparations. This approach was not feasible since chlamydiae are insensitive to the action of lysozyme (36) that is routinely used to prepare spheroplasts from which outer membranes are isolated after osmotic lysis. Also, the extremely small quantities of purified C. trachomatis organisms that can be practically obtained discouraged investigations directed at isolating outer membranes from mechanically disrupted organisms, since we felt that this method would result in very small yields of homogeneous membrane.…”

Section: Resultsmentioning

confidence: 99%

Purification and partial characterization of the major outer membrane protein of Chlamydia trachomatis

Caldwell¹,

Kromhout²,

Schachter³

1981

Infect Immun

993

473

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Elementary bodies (EB) of Chlamydia trachomatis serotypes C, E, and L2 were extrinsically radioiodinated, and whole-cell lysates of these serotypes were compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Autoradiography of the polypeptide profiles identified a major surface protein with an apparent subunit molecular weight of 39,500 that was common to each C. trachomatis serotype. The abilities of nonionic (Triton X-100), dipolar ionic (Zwittergent TM-314), mild (sodium deoxycholate and sodium N-lauroyl sarcosine), and strongly anionic (SDS) detergents to extract this protein from intact EB of the L2 serotype were investigated by SDS-PAGE analysis of the soluble and insoluble fractions obtained after each detergent treatment. Only SDS readily extracted this protein from intact EB. Sarkosyl treatment selectively solubilized the majority of other EB proteins, leaving the 39,500-dalton protein associated with the Sarkosyl-insoluble fraction. Ultrastructural studies of the Sarkosyl-insoluble EB pellet showed it to consist of empty EB particles possessing an apparently intact outer membrane. No structural evidence for a peptidoglycanlike cell wall was found. Morphologically these chlamydial outer membrane complexes (COMC) resembled intact chlamydial EB outer membranes. The 39,500-dalton outer membrane protein was quantitatively extracted from COMC by treating them with 2% SDS at 60°C. This protein accounted for 61% of the total COMC-associated protein, and its extraction resulted in a concomitant loss of the COMC membrane structure and morphology. The soluble extract obtained from SDS-treated COMC was adsorbed to a hydroxylapatite column and eluted with a linear sodium phosphate gradient. The 39,500-dalton protein was eluted from the column as a single peak at a phosphate concentration of approximately 0.3 M. The eluted protein was nearly homogeneous by SDS-PAGE and appeared free of contaminating carbohydrate, glycolipid, and nucleic acid. Hyperimmune mouse antiserum prepared against the 39,500-dalton protein from serotype L2 reacted with C. trachomatis serotypes Ba, E, D, K, Ll, L2, and L3 by indirect immunofluorescence with EB but failed to react with serotypes A, B, C, F, G, H, I, and J, with the C. trachomatis mouse pneumonitis strain, or with the C. psittaci feline pneumonitis, guinea pig inclusion conjunctivitis, or 6BC strains. Thus, the 39,500-dalton major outer membrane protein is a serogroup antigen of C. trachomatis organisms. Chlamydia are obligate intracellular bacteria These findings suggest both biochemical and characterized by a unique and complex growth structural similarities between chlamydial and cycle. Phylogenetically, the chlamydiae are gram-negative bacterial outer membranes. The closely related to gram-negative organisms. In outer membrane of gram-negative bacteria has particular, both gram-negative bacteria and been intensely investigated with respect to its chlamydiae have outer membranes that respond composition and function (4). Unique proteins, similarly to ...

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Section: Resultsmentioning

confidence: 99%

Purification and partial characterization of the major outer membrane protein of Chlamydia trachomatis

Caldwell¹,

Kromhout²,

Schachter³

1981

Infect Immun

993

473

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“…In L cells, NADP stimulates the multiplication of C. psittaci 6BC (142), and in host-free C. psittaci MN/Cal 10, it increases the release of CO2 from labeled glucose (506) and brings about the reduction of added cytochrome c (7,8,464). These effects of NADP deserve further investigation in light of the failure of many host-independent bacteria to transport NAD intact (for references, see reference 14).…”

Section: Synthesis Of Proteinmentioning

confidence: 99%

Interaction of chlamydiae and host cells in vitro.

Moulder¹

1991

Microbiological Reviews

513

288

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“…The cell surface structures, or envelopes, of these two forms have been reported to be similar, both morphologically and in cell wall amino acid analysis, to those of gram-negative bacteria such as Escherichia coli (15,24,27). However, the inability to detect muramic acid in purified EB or their cell walls (9,15) and the failure of lysozyme-ethylenediaminetetraacetate (EDTA) to effect the integrity of the envelopes of Chlamydia (25) indicate that the cell envelopes of Chlamydia differ fundamentally from those of most bacteria. Thus, the question arose as to whether the chlamydial cell surface structures maintain their organization in the same way as walls of gram-negative or gram-positive bacteria.…”

Section: Suspensions Of Isolated Cell Envelopes Of Infectious Elementmentioning

confidence: 99%

Effect of alkali on the structure of cell envelopes of Chlamydia psittaci elementary bodies

Narita¹,

Wyrick²,

Manire³

1976

J Bacteriol

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Suspensions of isolated cell envelopes of infectious elementary bodies (EB) of Chlamydia psittaci at alkaline pH showed a rapid, extensive decrease in absorbance, accompanied by the release of a cell envelope component in a sedimentable form. This phenomenon was observed both at 0 C and with envelopes which had been previously heated to 100 C. Monovalent and divalent cations effectively inhibited the turbidity loss, whereas ethylenediaminetetraacetate (EDTA) caused an accelerated decrease in turbidity. The turbidity loss observed after incubation of the envelopes at alkaline pH could be reversed to the level of the initial value by dialysis against distilled water containing Mg2+. Thin-section electron photomicrographs of purified EB exposed to alkaline buffer with EDTA revealed the loss of the internal contents of cells, but these cells still maintained their round shapes. The cell surface of treated EB appeared pitted in negatively stained preparations, whereas intact EB had a smooth surface. Electron microscopic studies on negatively stained preparations of the clear supernatant obtained after the treatment of the envelope with alkaline buffer containing EDTA demonstrated the presence of spherical particles, approximately 6 to 7 nm in diameter, and rodlike particles, which appeared to be made up of two or more spherical particles.

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Cytochrome C Reductase Activity of Meningopneumonitis Organisms at Different Stages of Development

Cited by 7 publications

References 2 publications

Purification and partial characterization of the major outer membrane protein of Chlamydia trachomatis

Purification and partial characterization of the major outer membrane protein of Chlamydia trachomatis

Interaction of chlamydiae and host cells in vitro.

Effect of alkali on the structure of cell envelopes of Chlamydia psittaci elementary bodies

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