Identification and Immunogenicity of Group A<i>Streptococcus</i>Culture Supernatant Proteins

Lei, Benfang; Mackie, Stacy M.; Łukomski, Sławomir; Musser, James M.

doi:10.1128/iai.68.12.6807-6818.2000

Cited by 142 publications

(154 citation statements)

References 54 publications

(56 reference statements)

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“…We postulate that increased expression of RsbU represents an attempt by S. pyogenes to activate the SigmaB stress response to cope with the effects of fluoride. Several examples are present in the literature of the application of proteomic analysis to the study of Streptococcus species (19,35,36). None of these studies examined the effect of fluoride on S. pyogenes and cannot be used to confirm our findings.…”

Section: Effect Of Fluoride On Protein Expression In S Pyogenes Cultcontrasting

confidence: 39%

Fluoride Exposure Attenuates Expression of Streptococcus pyogenes Virulence Factors

Thongboonkerd¹,

Luengpailin²,

Cao³

et al. 2002

Journal of Biological Chemistry

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Fluoridation causes an obvious reduction of dental caries by interference with cariogenic streptococci. However, the effect of fluoride on group A streptococci that causes rheumatic fever and acute poststreptococcal glomerulonephritis is not known. We have used proteomic analysis to create a reference proteome map for Streptococcus pyogenes and to determine fluoride-induced protein changes in the streptococci. Cellular and extracellular proteins were resolved by two-dimensional polyacrylamide gel electrophoresis and identified by matrix-assisted laser desorption ionization mass spectrometry. 183 protein spots were visualized, and 74 spots representing 60 unique proteins were identified. A 16-h exposure to sodium fluoride caused decreased expression of proteins required to respond to cellular stress, including anti-oxidants, glycolytic enzymes, transcriptional and translational regulators, and protein folding. Fluoride caused decreased cellular expression of two well-characterized S. pyogenes virulence factors. Fluoride decreased expression of glyceraldehyde-3-phosphate dehydrogenase, which acts to bind fibronectin and promote bacterial adherence. We also performed proteomic analysis of protein released by S. pyogenes into the culture supernatant and observed decreased expression of M proteins following fluoride exposure. These data provide evidence that fluoride causes decreased expression by S. pyogenes proteins used to respond to stress, virulence factors, and implicated in nonsuppurative complications of S. pyogenes, including glomerulonephritis and rheumatic fever.The incidence of some sequelae of group A streptococcal infection such as rheumatic fever and acute post-streptococcal glomerulonephritis (APSGN) 1 has decreased over the last five decades in the United States and western countries (1-3). However, these disorders continue unabated and are important public health problems in developing countries (1,3,4). The effects of fluoride on cariogenic Streptococcus mutans and other bacteria have been extensively studied. Fluoride inhibits enolase, a glycolytic enzyme (5, 6), inhibits F-ATPase activity resulting in less acidurance (7,8), reduces glucan-binding lectin activity (9), and decreases glucose incorporation (10). However, the effects of fluoride on group A Streptococci have not been examined. To examine simultaneous changes in multiple virulence factors, we performed a proteomic analysis of S. pyogenes exposed to fluoride. Western blotting and other immunological methods have been successfully used to study protein expression of various microorganisms, cells, and tissues. However, these techniques are constrained by the limited number of proteins that can be studied in each experiment and the availability of specific antibodies. Proteomic techniques date to 1975, when two-dimensional PAGE was simultaneously described by O'Farrell and Klose (11, 12) and applied to the study of a large number of proteins simultaneously. In twodimensional PAGE proteins are separated by differential isoelectric point (pI)...

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Section: Effect Of Fluoride On Protein Expression In S Pyogenes Cultcontrasting

confidence: 39%

Fluoride Exposure Attenuates Expression of Streptococcus pyogenes Virulence Factors

Thongboonkerd¹,

Luengpailin²,

Cao³

et al. 2002

Journal of Biological Chemistry

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“…Assessment of in Vitro Production of SlaA-GAS strains used to test for in vitro production of SlaA were cultured in protein-reduced ToddHewitt broth containing 0.2% (w/v) yeast extract (PR-THY; Difco Laboratories, Detroit, MI) in 5% CO 2 at 37°C (20). Bacteria were grown to mid-exponential, early stationary phase, and overnight, centrifuged, and the pellet was discarded.…”

Section: Methodsmentioning

confidence: 99%

Analysis of a Novel Prophage-encoded Group A Streptococcus Extracellular Phospholipase A2

Nagiec

Lei

Parker

et al. 2004

Journal of Biological Chemistry

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Group A Streptococcus (GAS) is an important human pathogen that causes many types of infections, including pharyngitis and severe invasive diseases. We recently sequenced the genome of a serotype M3 strain and identified a prophage-encoded secreted phospholipase A 2 designated SlaA. To study SlaA structure-activity relationships, 20 site-specific mutants were constructed by alanine-replacement mutagenesis and purified to apparent homogeneity. Enzymatic activity was greatly reduced by alanine replacement of amino acid residues previously described as crucial in the catalytic mechanism of secreted phospholipase A 2 . Similarly, substitution of five residues in an inferred Ca 2؉ -binding loop and three residues in the inferred active site region resulted in loss of activity of 76.5% or greater relative to the wild-type enzyme. Analysis of enzyme substrate specificity confirmed SlaA as a phospholipase A 2 , with activity against multiple phospholipid head groups and acyl chains located at the sn-2 position. PCR analysis of 1,189 GAS strains representing 48 M protein serotypes commonly causing human infections identified the slaA gene in 129 strains of nine serotypes (M1, M2, M3, M4, M6, M22, M28, M75, and st3757). Expression of SlaA by strains of these serotypes was confirmed by Western immunoblot. SlaA production increased rapidly and substantially on co-culture with Detroit 562 human pharyngeal epithelial cells. Together, these data provide new information about a novel extracellular enzyme that participates in GAS-human interactions.

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“…These authors also demonstrated considerable cross-protection of the 30-valent vaccine against non-vaccine M serotypes [4]. Recently, a series of independent studies have documented a class of GAS cell wall-associated or secreted metabolic enzymes that contain neither N-terminal leader sequences nor C-terminal cell wall anchors [5][6][7][8][9]. Perhaps because these molecules do not fit common categories of bacterial vaccine antigen, e.g.…”

mentioning

confidence: 99%

Conserved anchorless surface proteins as group A streptococcal vaccine candidates

et al. 2012

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Identification and Immunogenicity of Group AStreptococcusCulture Supernatant Proteins

Cited by 142 publications

References 54 publications

Fluoride Exposure Attenuates Expression of Streptococcus pyogenes Virulence Factors

Fluoride Exposure Attenuates Expression of Streptococcus pyogenes Virulence Factors

Analysis of a Novel Prophage-encoded Group A Streptococcus Extracellular Phospholipase A2

Conserved anchorless surface proteins as group A streptococcal vaccine candidates

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