Production of Group C Streptococcus Phage-associated Lysin and the Preparation of Streptococcus pyogenes Protoplast Membranes

Wheeler, J. W.; Holland, J P; Terry, J. M.; Blainey, J. D.

doi:10.1099/00221287-120-1-27

Cited by 5 publications

(4 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Consequently, this enzyme has been used as a molecular tool for decades to isolate cell-wall-linked proteins and extract DNA from group A streptococci (9,10). More recently, we have shown that the bacteriolytic properties of PlyC can protect mice from streptococcal challenge, suggesting a therapeutic use of the enzyme (11).…”

mentioning

confidence: 99%

PlyC: A multimeric bacteriophage lysin

Nelson

Schuch

Chahales

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

190

189

View full text Add to dashboard Cite

Lysins are murein hydrolases produced by bacteriophage that act on the bacterial host cell wall to release progeny phage. When added extrinsically in their purified form, these enzymes produce total lysis of susceptible Gram-positive bacteria within seconds, suggesting a unique antimicrobial strategy. All known Grampositive lysins are produced as a single polypeptide containing a catalytic activity domain, which cleaves one of the four major peptidoglycan bonds, and a cell-wall-binding domain, which may bind a species-specific carbohydrate epitope in the cell wall. Here, we have cloned and expressed a unique lysin from the streptococcal bacteriophage C1, termed PlyC. Molecular characterization of the plyC operon reveals that PlyC is, surprisingly, composed of two separate gene products, PlyCA and PlyCB. Based on biochemical and biophysical studies, the catalytically active PlyC holoenzyme is composed of eight PlyCB subunits for each PlyCA. Inhibitor studies predicted the presence of an active-site cysteine, and bioinformatic analysis revealed a cysteine, histidine-dependent amidohydrolase/peptidase domain within PlyCA. Point mutagenesis confirmed that PlyCA is responsible for the observed catalytic activity, and Cys-333 and His-420 are the active-site residues. PlyCB was found to self-assemble into an octamer, and this complex alone was able to direct streptococcal cell-wall-specific binding. Similar to no other proteins in sequence databases, PlyC defines a previously uncharacterized structural family of cell-wall hydrolases.multimeric protein ͉ cell-wall hydrolase ͉ Streptococcus

show abstract

mentioning

confidence: 99%

PlyC: A multimeric bacteriophage lysin

Nelson

Schuch

Chahales

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

190

189

View full text Add to dashboard Cite

show abstract

“…C 1 phage lysin can cause ''lysis from without'' in groups A and E as well as group C streptococci (12,13). This unique activity has been exploited as a tool in group A streptococcal studies to isolate surface molecules including M proteins (14), to lyse cells for DNA extraction, and to make protoplasts when used in a hypertonic environment (15).…”

mentioning

confidence: 99%

Prevention and elimination of upper respiratory colonization of mice by group A streptococci by using a bacteriophage lytic enzyme

Nelson

Loomis²,

Fischetti³

2001

Proc. Natl. Acad. Sci. U.S.A.

481

415

View full text Add to dashboard Cite

Bacteriophage lytic enzymes quickly destroy the cell wall of the host bacterium to release progeny phage. Because such lytic enzymes specifically kill the species in which they were produced, they may represent an effective way to control pathogenic bacteria without disturbing normal microflora. In this report, we studied a murein hydrolase from the streptococcal bacteriophage C 1 termed lysin. This enzyme is specific for groups A, C, and E streptococci, with little or no activity toward several oral streptococci or other commensal organisms tested. Using purified lysin in vitro, we show that 1,000 units (10 ng) of enzyme is sufficient to sterilize a culture of Ϸ10 7 group A streptococci within 5 seconds. When a single dose of lysin (250 units) is first added to the oral cavity of mice, followed by 10 7 live group A streptococci, it provides protection from colonization (28.5% infected, n ‫؍‬ 21) compared with controls without lysin (70.5% infected, n ‫؍‬ 17) (P < 0.03). Furthermore, when lysin (500 units) was given orally to 9 heavily colonized mice, no detectable streptococci were observed 2 h after lysin treatment. In all, these studies show that lysin represents a unique murein hydrolase that has a rapid lethal effect both in vitro and in vivo on group A streptococci, without affecting other indigenous microorganisms analyzed. This general approach may be used to either eliminate or reduce streptococci from the upper respiratory mucosal epithelium of either carriers or infected individuals, thus reducing associated disease.

show abstract

“…Preparation of SCM. SCM was prepared from Streptococcus pyogenes (T type 12, Tanaka strain), isolated from the throat of a patient with APSGN, by a modification of the method of Wheeler et al (19) with group C streptococcus bacteriophage-associated lysin, as described previously (20). The SCM obtained had a rhamnose concentration of less than 0.5% (6).…”

Section: Methodsmentioning

confidence: 99%

High levels of antibodies to streptococcal cell membrane antigens specifically bound to monoclonal antibodies in acute poststreptococcal glomerulonephritis

Yoshimoto¹,

Hosoi²,

Fujisawa³

et al. 1987

J Clin Microbiol

View full text Add to dashboard Cite

We produced 15 immunoglobulin G class monoclonal antibodies against antigens of the streptococcal cell membrane (SCM) of Streptococcus pyogenes (T type 12, Tanaka strain) and determined the levels in human sera of antibodies against Triton-X-extracted antigens specifically bound to each of these 15 monoclonal antibodies by enzyme-linked immunosorbent assay. Sample sera were obtained from 10 normal controls (group 1), 10 patients with streptococcal pharyngitis without sequelae (group 2), and 8 patients with acute poststreptococcal glomerulonephritis (APSGN) (group 3). Anti-streptolysin O (ASO) titers of the sera increased in the order of groups 1, 2, and 3. There was no relationship between ASO titer and the level of anti-SCM antibodies, and there was no significant difference in the level of anti-SCM antibodies determined with each of the 15 monoclonal antibodies between group 1 and group 2 sera. Group 3 sera had higher levels of antibodies to SCM antigens specifically bound to each of 14 of these 15 monoclonal antibodies than group 1 or group 2 sera did. Of these 14 monoclonal antibodies, 9 reacted with the four SCM antigens separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretically transferred to a nitrocellulose sheet. These results suggest that high levels of antibodies to SCM antigens are related to the development of APSGN and that the systemic immune response to SCM antigens is involved in the pathogenesis of APSGN.

show abstract

Production of Group C Streptococcus Phage-associated Lysin and the Preparation of Streptococcus pyogenes Protoplast Membranes

Cited by 5 publications

References 14 publications

PlyC: A multimeric bacteriophage lysin

PlyC: A multimeric bacteriophage lysin

Prevention and elimination of upper respiratory colonization of mice by group A streptococci by using a bacteriophage lytic enzyme

High levels of antibodies to streptococcal cell membrane antigens specifically bound to monoclonal antibodies in acute poststreptococcal glomerulonephritis

Contact Info

Product

Resources

About