Localization of Enzymes in the Microbial Cell

Alexander, Martin

doi:10.1128/mmbr.20.2.67-93.1956

Cited by 35 publications

(22 citation statements)

References 8 publications

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“…VOL. 1,1970 on August 1, 2020 by guest http://iai.asm.org/ Downloaded from DISCUSSION From the results obtained here and in the accompanying report (7), it is apparent that the protective antigen of E. rhusiopathiae found in culture supernatant fluids exists in a variety of conditions, ranging from material that cannot be sedimented by centrifugation at 198,000 X g for 12 hr to aggregates that sediment at much lower centrifugal speeds. In these studies, the aggregated material was selected for further use in purification because protective antigen activity, existing as a readily sedimented particle, offered a fortuitous way of separating the protective antigen from the wide diversity of soluble horse serum proteins which constituted a major portion of the original medium.…”

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confidence: 55%

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Solubilization and Characterization of a Protective Antigen of Erysipelothrix rhusiopathiae

White¹,

Verwey

1970

Infect Immun

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A particulate fraction of Erysipelothrix rhusiopathiae cultures has been subjected to butanol extraction and treatment with surface-active agents in an attempt to solubilize a protective antigen. The particulate fraction was partitioned into the butanol layer but was not solubilized. Only sodium dodecyl sulfate solubilized the particles. The soluble protective activity was not sedimented by centrifugation at 198,000 × g for 12 hr but was excluded by Sephadex G-200. Immunodiffusion studies of the soluble fraction demonstrated eight antigens, five of horse serum origin and three of E. rhusiopathiae origin. Ultracentrifugation indicated that spontaneous reaggregation occurred after removal of the sodium dodecyl sulfate. Analytical ultracentrifugation showed that 90% of the sodium dodecyl sulfatetreated material migrated as a single homogeneous 3.5 S component. The physical and biological characteristics of the protective activity suggest that the protective antigen is a glyco-lipoprotein.

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confidence: 55%

“…All preparations were then sequentially centrifuged in a Spinco model L ultracentrifuge, and the fractions obtained were tested for protective activity. The fractions were named by use of the system of Alexander (1). Thus, 20P and 20S represented the sediment and supernatant fluid, respectively, obtained after centrifugation at 20,000 X g.…”

Section: Methodsmentioning

confidence: 99%

Solubilization and Characterization of a Protective Antigen of Erysipelothrix rhusiopathiae

White¹,

Verwey

1970

Infect Immun

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“…After a final centrifugation of 140,000 X g for 12 hr, the sediments were each resuspended to a volume equal to that of the supernatant fluid from which they were removed, and these, along with the final supernatant fluid, were tested for protective activity. The sediments were designated the lOP 36P, 81P, llOP, 140P, and 140P-12-hr fractions, and the final supernatant fluid was designated the 140S fraction by using the system suggested by Alexander (1).…”

Section: Resultsmentioning

confidence: 99%

Isolation and Characterization of a Protective Antigen-Containing Particle from Culture Supernatant Fluids of Erysipelothrix rhusiopathiae

White¹,

Verwey

1970

Infect Immun

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The mouse-protective activity of Erysipelothrix rhusiopathiae culture supernatant fluids exists in a polydisperse form, ranging in density from aggregates which sediment at 10,000 × g for 3 hr to soluble units which will not sediment at 198,000 × g for 12 hr. A partially purified protective antigen has been isolated from the aggregates sedimented from a concentrate of the culture supernatant fluid at 20,000 × g for 3 hr. These aggregates contained the major protective antigen or antigens of E. rhusiopathiae , since, in addition to inducing active immunity, they adsorbed essentially all of the passively protecting antibody from rabbit antiserum produced by immunization with whole culture. The protective activity in these aggregates was destroyed by trypsin and greatly diminished by muramidase and heating at 64 C, but was not affected by lipase or ribonuclease.

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“…The centrifuge fractions are designated by the terminology suggested by Alexander (12). The letter p or s refers to precipitate or supernate, the prefix indicates the force in 1,000 G and the suffix refers to time in minutes.…”

Section: Chemical Analyses--nitrogen Was Determined By the Micro-kjementioning

confidence: 99%

Identification of a Toxic Cellular Component of Group a Streptococci as a Complex of Group-Specific C Polysaccharide and a Protein

Schwab

Cromartie

Roberson

1959

The Journal of Experimental Medicine

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Chronic multinodular skin lesions were produced in rabbits injected intradermally at a single site with a sterile extract of sonically disrupted Group A streptococcal cells (1). These lesions, which consisted of separate nodules scattered over areas as large as 12 cm. in diameter, presented several remarkable features. The injury to dermal connective tissue was evident within 48 hours after injection, suggesting that the material had a primary toxic effect. The reaction was observed to continue for as long as 30 days in the first series of experiments (1), and subsequent studies have revealed it to continue for at least 60 days after a single injection.In the gross, the process was characterized by periods of abatement followed by either exacerbation of the subsiding lesions or the development of additional nodules in areas where the lesions have previously completely subsided. The initial nodules and the recurrent nodular lesions were associated with focal necrosis of dermal connective tissue. What appeared to be acute focal necrosis was noted during the first 48 hours after injection and as late as 60 days after the animals were inoculated. As the process evolved, mononuclear and giant cells were found in the tissues along with the areas of acute focal necrosis.The gross and microscopic changes suggested that a single injection of this sterile extract could initiate a mechanism of injury that caused recurrent acute focal damage of connective tissue extending over a period of at least 60 days. Comparative study of extracts of various species of streptococci suggested that this toxic material was a cellular component characteristic of Group A streptococci (2). I t is the purpose of this report to record data associating the toxic activity with a complex macromolecule containing the Group-specific C polysaccharide. Materials and MethodsExtraction of Cells.--The cultivation, harvesting, and extraction of the organisms has been described (1). The cells from an 18 hour broth culture were washed 2 times with cold saline

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Localization of Enzymes in the Microbial Cell

Abstract: A wealth of literature exists on the morphology Schneider and their co-workers (47), and the and cytological relationships of the internal approach more recently has been extended to structures of the microbial cell, but the organizaplant tissue with equally fruitful results (37).

Cited by 35 publications

References 8 publications

Solubilization and Characterization of a Protective Antigen of Erysipelothrix rhusiopathiae

Solubilization and Characterization of a Protective Antigen of Erysipelothrix rhusiopathiae

Isolation and Characterization of a Protective Antigen-Containing Particle from Culture Supernatant Fluids of Erysipelothrix rhusiopathiae

Identification of a Toxic Cellular Component of Group a Streptococci as a Complex of Group-Specific C Polysaccharide and a Protein

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