Detection of Agr-Type Autoinducing Peptides Produced by Staphylococcus aureus

2017 ◽  
pp. 89-96
Author(s):  
Ewan J. Murray ◽  
Paul Williams
Keyword(s):  
Staphylococcus Aureus ◽  
Autoinducing Peptides

scholarly journals Characterization of structural elements in native autoinducing peptides and non-native analogues that permit the differential modulation of AgrC-type quorum sensing receptors in Staphylococcus aureus

2016 ◽  
Vol 14 (1) ◽  
pp. 113-121 ◽  
Author(s):  
Yftah Tal-Gan ◽  
Monika Ivancic ◽  
Gabriel Cornilescu ◽  
Helen E. Blackwell
Keyword(s):  
Staphylococcus Aureus ◽  
Quorum Sensing ◽  
Structural Elements ◽  
Differential Modulation ◽  
Autoinducing Peptides

Structural analyses of autoinducing peptides and analogues thereof reveal motifs critical for modulation of quorum sensing receptors inS. aureus.

scholarly journals Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein

2007 ◽  
Vol 73 (19) ◽  
pp. 6036-6044 ◽  
Author(s):  
Cheryl L. Malone ◽  
Blaise R. Boles ◽  
Alexander R. Horswill
Keyword(s):  
Staphylococcus Aureus ◽  
Fusion Protein ◽  
Biochemical Analysis ◽  
Side Chain ◽  
Biological Applications ◽  
Dna Encoding ◽  
Sensing System ◽  
Quorum Sensing System ◽  
Reporter Strains ◽  
Autoinducing Peptides

ABSTRACT The Agr quorum-sensing system of Staphylococcus aureus modulates the expression of virulence factors in response to autoinducing peptides (AIPs). The peptides are seven to nine residues in length and have the C-terminal five residues constrained in a thiolactone ring. We have developed a new method to generate AIP structures using an engineered DnaB mini-intein from Synechocystis sp. strain PCC6803. In the method, an oligonucleotide encoding the AIP is ligated to the intein and the fusion protein is expressed and purified by affinity chromatography. To produce the correct AIP structure, intein splicing is interrupted, allowing the cysteine side chain to catalyze thiolactone ring formation and release AIP from the resin. The technique is simple and robust, and we have successfully produced the three main classes of AIPs using the intein system. The intein-generated AIPs possessed the correct thiolactone ring modification based on biochemical analysis, and, importantly, all the samples were bioactive against S. aureus. The AIP activity was confirmed through Agr interference and activation profiling with developed S. aureus reporter strains. The simplicity of the method, benefits of DNA encoding, and scalable nature enable the production of S. aureus AIPs for many biological applications.

scholarly journals Gaussia Luciferase as a Reporter for Quorum Sensing in Staphylococcus aureus

Sensors ◽  
2020 ◽  
Vol 20 (15) ◽  
pp. 4305
Author(s):  
Isobel Blower ◽  
Carmen Tong ◽  
Xiaohui Sun ◽  
Ewan Murray ◽  
Jeni Luckett ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Quorum Sensing ◽  
Quantitative Assessment ◽  
Chemical Agent ◽  
Secreted Protein ◽  
Human Pathogen ◽  
Gaussia Luciferase ◽  
Bacterial Pathogenicity ◽  
Autoinducing Peptides

Gaussia luciferase (GLuc) is a secreted protein with significant potential for use as a reporter of gene expression in bacterial pathogenicity studies. To date there are relatively few examples of its use in bacteriology. In this study we show that GLuc can be functionally expressed in the human pathogen Staphylococcus aureus and furthermore show that it can be used as a biosensor for the agr quorum sensing (QS) system which employs autoinducing peptides to control virulence. GLuc was linked to the P3 promoter of the S. aureusagr operon. Biosensor strains were validated by evaluation of chemical agent-mediated activation and inhibition of agr. Use of GLuc enabled quantitative assessment of agr activity. This demonstrates the utility of Gaussia luciferase for in vitro monitoring of agr activation and inhibition.

Cytochemical Studies of Cell Wall Biosynthesis in Staphylococcus Aureus

1972 ◽  
Vol 30 ◽  
pp. 328-329
Author(s):  
Masaatsu Koike ◽  
Koichi Nakashima ◽  
Kyoko Iida
Keyword(s):  
Staphylococcus Aureus ◽  
Periodate Oxidation ◽  
Early Time ◽  
The Other ◽  
Penicillin G ◽  
Cell Wall Biosynthesis ◽  
Septal Wall ◽  
Peptidoglycan Biosynthesis ◽  
Gram Positive Bacteria ◽  
Cross Linkage

Penicillin exerts the activity to inhibit the peptide cross linkage between each polysaccharide backbone at the final stage of wall-peptidoglycan biosynthesis of bacteria. Morphologically, alterations of the septal wall and mesosome in gram-positive bacteria, which were occurred in early time after treatment with penicillin, have been observed. In this experiment, these alterations were cytochemically investigated by means of silver-methenamine staining after periodate oxidation, which is applied for detection of localization of wall mucopolysaccharide.Staphylococcus aureus strain 209P treated with 100 u/ml of penicillin G was divided into two aliquotes. One was fixed by Kellenberger-Ryter's OSO4 fixative at 30, 60 and 120 min after addition of the antibiotic, dehydrated through alcohol series, and embedded in Epon 812 (Specimen A). The other was fixed by 21 glutaraldehyde, dehydrated through glycolmethacrylate series and embedded in glycolmethacrylate mixture, according to Bernhard's method (Specimen B).

Gold labeling of teichoic acid on adjacent surfaces of staphylococcus aureus

1990 ◽  
Vol 48 (3) ◽  
pp. 578-579
Author(s):  
Margaret Hukee
Keyword(s):  
Staphylococcus Aureus ◽  
Teichoic Acid ◽  
Protein Labeling ◽  
Minimal Amount ◽  
Section Thickness ◽  
Double Labeling ◽  
Parallel Surfaces ◽  
Supporting Membrane

Gold labeling of two antigens (double labeling) is often done on two section surfaces separated by section thickness. Whether labeling is done on both sides of the same section or on two parallel surfaces separated by section thickness (PSSST), comparable results are dependent on an equal number of epitopes being exposed at each surface. We propose a method to study protein labeling within the same field of proteins, by examining two directly adjacent surfaces that were split during sectioning. The number of labeling sites on adjacent surfaces (AS) were compared to sites on PSSST surfaces in individual bacteria.Since each bacteria needed to be recognizable in all three section surfaces, one-hole grids were used for labeling. One-hole grids require a supporting membrane and excessive handling during labeling often ruptures the membrane. To minimize handling, a labeling chamber was designed that is inexpensive, disposable, minimizes contamination, and uses a minimal amount of solution.

Studies on the Relationship of Staphylococcus Aureus to Pseudomembranous Enteritis and to Postantibiotic Enteritis

1960 ◽  
Vol 38 (3) ◽  
pp. 441-451 ◽  
Author(s):  
William H. Dearing ◽  
Archie H. Baggenstoss ◽  
Lyle A. Weed
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomembranous Enteritis ◽  
Relationship Of ◽  
The Relationship
Sign in / Sign up

Export Citation Format

Share Document