Comparative Electrophoretic and Amino Acid Analyses of Isolated Membranes from
            <i>Streptococcus pyogenes</i>
            and Stabilized L-Form

Panos, Charles; Fagan, G.; Zarkadas, Constantinos G.

doi:10.1128/jb.112.1.285-290.1972

Cited by 9 publications

(6 citation statements)

References 13 publications

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“…Chemical fractionation of Sig-1 membranes gives no evidence of muramic acid at a detection limit of 6 umol/g (dry weight) of membrane. Hydrolyzed membranes contain 0.8% hexosamine, which is similar to that found in hydrolysates of membranes from B. subtilis protoplasts (0.6%) (1), Acholeplasma (0.5%) (12), and an L-form of Streptococcus pyogenes (1.2%) (11). The characterization of this hexosamine is in progress.…”

Section: Discussionsupporting

confidence: 60%

Adaptation of a stable L-form of Bacillus subtilis to minimal salts medium without osmotic stabilizers

Gilpin¹,

Patterson²

1976

J Bacteriol

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An L-form isolated from Bacillus subtilis 168 was adapted to growth in a 340 mOsm minimal salts medium without the addition of osmotically protective solutes. This L-form had no chemically detectable peptidoglycan residues on its surface, but 0.8% of the dry weight of washed membranes was hexosamine. The osmotic stability and susceptibility to bacitracin and vancomycin of the L-form adapted to growth in 340 mOsm osmotically unprotected medium was twice that of the L-form grown in 2,680 mOsm medium supplemented with 1.2 M NaCl.

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Section: Discussionsupporting

confidence: 60%

Adaptation of a stable L-form of Bacillus subtilis to minimal salts medium without osmotic stabilizers

Gilpin¹,

Patterson²

1976

J Bacteriol

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“…The total protein and amino acid composition of the membranes from both type 6 and type 49 streptococci did not vary between phases of growth, and the mole ratios of amino acids compared well with those previously published (7,19). Cysteine and tryptophan were undetectable in our membrane preparations.…”

Section: Pyogenessupporting

confidence: 88%

Chemical Analysis of Changes in Membrane Composition During Growth of Streptococcus pyogenes

Rijn

Kessler

1979

Infect Immun

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Changes in the structural components ofthe Streptococcuspyogenes membrane between exponential and early stationary phases of growth are reported. The overall protein composition ranged from 70 to 73% of the dry weight of the membranes, irrespective of the phase of growth from which they were isolated. Amino acid analyses of membranes isolated from streptococci in either the exponential or stationary phase of growth demonstrated that two amino acids, cysteine and tryptophan, were absent. Further analysis of the membrane proteins by sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis demonstrated that there were proteins unique to a particular phase of growth as well as differences in the amount of specific proteins from the various growth phases. In addition, membranes isolated from exponential-phase cultures contained a higher percentage of peripheral protein than did stationary-phase membranes. There also appeared to be an increase in the amount of outer surface proteins during this growth phase. The phosphorus content of the membranes increased during the stationary phase of growth, whereas the sugar composition remained constant. The only sugar found under various conditions of growth in any of the strains was glucose. Total fatty acid content and the mole percent composition of various fatty acids did not change in the different phases of growth. However, the mole percent composition of fatty acids in the membranes of various group A streptococci did differ between strains. Therefore, these results provide evidence that the composition of membranes of S. pyogenes does not remain constant throughout the growth phases of the culture.

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“…As already indicated, it is also possible that this lack of D-alanine in the TA from the L-form may be due to membrane structural or conformational changes after conversion from the streptococcus. Disc gel electrophoretic studies have already presented evidence for the existence of marked quantitative differences in the membrane protein patterns of this S. pyogenes and its L-form (26).…”

Section: Discussionmentioning

confidence: 95%

Teichoic Acid of a Stabilized L-Form ofStreptococcus pyogenes

Slabyj

Panos

1973

J Bacteriol

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A stabilized L-form of Streptococcus pyogenes continues to synthesize glycerol teichoic acid. This polymer was obtained from S. pyogenes and its L-form, treated in identical fashion, and compared. Highly purified glycerol teichoic acid from only the L-form was found to be devoid of D-alanine and to have a shorter chain length. Otherwise, the glycerol teichoic acid from these two organisms was found to be a 1, 3-phosphodiester-linked glycerophosphate polymer substituted with D-glucose. Evidence is presented that most, if not all, of the glycerol teichoic acid in.this streptococcus lies between the wall and membrane. A possible need for the continued synthesis of a minute amount of glycerol teichoic acid by this L-form for survival is discussed in terms of the known function of teichoic acids in bacteria.MATERIALS AND METHODS Bacteria, media, and cell fractionation. S. pyogenes, type 12, and its stabilized L-form were used (7). S. pyogenes (5 to 8 liters) was incubated at 37 C for 16 h and harvested when in late stationary phase by centrifugation (17,000 x g) at 4 C. The streptococcus was cultured in brucella broth (Pfizer Diagnostics, Brooklyn, N.Y.) containing either bovine serum albumin, fraction V (8 g/liter; Armour Pharmaceutical Co., Chicago, Ill.) or supplemented with (in grams per liter): yeast extract, 8; sodium acetate, 10; glucose, 19; and KH2P04, 4.5, with the pH adjusted to 7.3. L-form cells were cultured in brucella broth containing bovine serum albumin with the addition of 3% NaCl (7) with and without penicillin G (1,000 U/ml). Where indicated, L-form and coccal media were supplemented, except that yeast extract was omitted. Supplemented medium readily caramelized when autoclaved. Thus, precautions had to be taken (i.e., rapid cooling after autoclaving) to prevent excessive caramelization. An alternative procedure, referred to as noncaramelized medium, denotes the addition of the supplement to the autoclaved basal medium after sterilization by membrane filtration (Millipore Corp.; 934 on July 31, 2020 by guest

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Comparative Electrophoretic and Amino Acid Analyses of Isolated Membranes from Streptococcus pyogenes and Stabilized L-Form

Cited by 9 publications

References 13 publications

Adaptation of a stable L-form of Bacillus subtilis to minimal salts medium without osmotic stabilizers

Adaptation of a stable L-form of Bacillus subtilis to minimal salts medium without osmotic stabilizers

Chemical Analysis of Changes in Membrane Composition During Growth of Streptococcus pyogenes

Teichoic Acid of a Stabilized L-Form ofStreptococcus pyogenes

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