scholarly journals Epitope Recognition of a Monoclonal Antibody Raised against a Synthetic Glycerol Phosphate Based Teichoic Acid

2021 ◽  
Author(s):  
Francesca Berni ◽  
Ermioni Kalfopoulou ◽  
Ana M. Gimeno Cardells ◽  
Filippo Carboni ◽  
Daan van der Es ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Teichoic Acid ◽  
Glycerol Phosphate ◽  
Epitope Recognition

scholarly journals Surface Glycopolymers Are Crucial forIn VitroAnti-Wall Teichoic Acid IgG-Mediated Complement Activation and Opsonophagocytosis of Staphylococcus aureus

2015 ◽  
Vol 83 (11) ◽  
pp. 4247-4255 ◽  
Author(s):  
Jong-Ho Lee ◽  
Na-Hyang Kim ◽  
Volker Winstel ◽  
Kenji Kurokawa ◽  
Jesper Larsen ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Teichoic Acid ◽  
Complement C3 ◽  
Glycerol Phosphate ◽  
Glycosylation Pattern ◽  
Wall Teichoic Acid ◽  
Content Type ◽  
Gram Positive Bacteria ◽  
Capsule Production ◽  
Cell Envelopes

ABSTRACTThe cell envelopes of many Gram-positive bacteria contain wall teichoic acids (WTAs).Staphylococcus aureusWTAs are composed of ribitol phosphate (RboP) or glycerol phosphate (GroP) backbones substituted withd-alanine andN-acetyl-d-glucosamine (GlcNAc) orN-acetyl-d-galactosamine (GalNAc). Two WTA glycosyltransferases, TarM and TarS, are responsible for modifying the RboP WTA with α-GlcNAc and β-GlcNAc, respectively. We recently reported that purified human serum anti-WTA IgG specifically recognizes β-GlcNAc of the staphylococcal RboP WTA and then facilitates complement C3 deposition and opsonophagocytosis ofS. aureuslaboratory strains. This prompted us to examine whether anti-WTA IgG can induce C3 deposition on a diverse set of clinicalS. aureusisolates. To this end, we compared anti-WTA IgG-mediated C3 deposition and opsonophagocytosis abilities using 13 different staphylococcal strains. Of note, the majority ofS. aureusstrains tested was recognized by anti-WTA IgG, resulting in C3 deposition and opsonophagocytosis. A minority of strains was not recognized by anti-WTA IgG, which correlated with either extensive capsule production or an alteration in the WTA glycosylation pattern. Our results demonstrate that the presence of WTAs with TarS-mediated glycosylation with β-GlcNAc in clinically isolatedS. aureusstrains is an important factor for induction of anti-WTA IgG-mediated C3 deposition and opsonophagocytosis.

scholarly journals Use of CDP-Glycerol as an Alternate Acceptor for the Teichoic Acid Polymerase Reveals that Membrane Association Regulates Polymer Length

2008 ◽  
Vol 190 (21) ◽  
pp. 6940-6947 ◽  
Author(s):  
Jeffrey W. Schertzer ◽  
Eric D. Brown
Keyword(s):  
Isotope Labeling ◽  
Teichoic Acid ◽  
Extracellular Polysaccharide ◽  
Membrane System ◽  
Polymer Synthesis ◽  
Glycerol Phosphate ◽  
Polysaccharide Biosynthesis ◽  
Steady State Kinetics ◽  
Length Regulation ◽  
Lag Period

ABSTRACT The study of bacterial extracellular polysaccharide biosynthesis is hampered by the fact that these molecules are synthesized on membrane-resident carrier lipids. To get around this problem, a practical solution has been to synthesize soluble lipid analogs and study the biosynthetic enzymes using a soluble system. This has been done for the Bacillus subtilis teichoic acid polymerase, TagF, although several aspects of catalysis were inconsistent with the results obtained with reconstituted membrane systems or physiological observations. In this work we explored the acceptor substrate promiscuity and polymer length disregulation that appear to be characteristic of TagF activity away from biological membranes. Using isotope labeling, steady-state kinetics, and chemical lability studies, we demonstrated that the enzyme can synthesize poly(glycerol phosphate) teichoic acid using the elongation substrate CDP-glycerol as an acceptor. This suggests that substrate specificity is relaxed in the region distal to the glycerol phosphate moiety in the acceptor molecule under these conditions. Polymer synthesis proceeded at a rate (27 min−1) comparable to that in the reconstituted membrane system after a distinct lag period which likely represented slower initiation on the unnatural CDP-glycerol acceptor. We confirmed that polymer length became disregulated in the soluble system as the polymers synthesized on CDP-glycerol acceptors were much larger than the polymers synthesized on the membrane or previously found attached to bacterial cell walls. Finally, polymer synthesis on protease-treated membranes suggested that proper length regulation is retained in the absence of accessory proteins and provided evidence that such regulation is conferred through proper association of the polymerase with the membrane.

Poly-N-acetylglucosamine and poly(glycerol phosphate) teichoic acid identification from staphylococcal biofilm extracts using excitation sculptured TOCSY NMR

2008 ◽  
Vol 4 (2) ◽  
pp. 170-174 ◽  
Author(s):  
Jane L. Wagstaff ◽  
Irina Sadovskaya ◽  
Evgeny Vinogradov ◽  
Saïd Jabbouri ◽  
Mark J. Howard
Keyword(s):  
Teichoic Acid ◽  
Glycerol Phosphate

Myc-epitope tagged proteins detected with the 9E10 antibody in immunofluorescence and immunoprecipitation assays but not in Western blot analysis

1998 ◽  
Vol 76 (1) ◽  
pp. 125-128 ◽  
Author(s):  
Huizhou Fan ◽  
Cristy Villegas ◽  
Arthur K Chan ◽  
Jim A Wright
Keyword(s):  
Gene Expression ◽  
Monoclonal Antibody ◽  
Recombinant Protein ◽  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Immunofluorescence Assay ◽  
Expression Vectors ◽  
Epitope Recognition ◽  
In Cells

A human Myc epitope is frequently used to tag proteins for expression experiments in nonhuman cells. We used the monoclonal 9E10 antibody specific for this epitope to analyse the expression of four proteins carrying the Myc tag in cells transfected with expression vectors. While all four proteins can be detected by immunofluorescence and immunoprecipitation assays, surprisingly, only two proteins could be detected in Western blot analysis, indicating that epitope recognition by the monoclonal antibody can be blocked in some membrane-retained ectopic proteins. Other techniques such as immunofluorescence and immunoprecipitation assays can be successfully used with the 9E10 antibody to determine potential expression of Myc-tagged proteins.Key words: recombinant protein, Myc epitope, 9E10, Western blot, gene expression, immunofluorescence assay, immunoprecipitation.

scholarly journals The wall content and composition of Bacillus subtilis var. niger grown in a chemostat

1970 ◽  
Vol 118 (3) ◽  
pp. 367-373 ◽  
Author(s):  
D. C. Ellwood
Keyword(s):  
Bacillus Subtilis ◽  
Phosphorus Content ◽  
Teichoic Acid ◽  
Dry Weight ◽  
Growth Conditions ◽  
Glycerol Phosphate ◽  
Limited Growth ◽  
Anionic Polymer ◽  
Rate Analysis ◽  
Teichuronic Acid

Bacillus subtilis var. niger was grown in a chemostat with various growth limitations and at various growth rates. The wall content and composition of the organism grown under these conditions were determined. The wall content, expressed as a percentage of the dry weight of organisms, varied with the growth rate. Analysis of wall samples showed that their composition also varied, particularly with respect to the phosphorus content. Wall samples extracted with trichloroacetic acid under carefully controlled conditions were found to contain various amounts of phosphorus, this being present as a glycerol phosphate polymer containing hexose (glucose and in some cases galactose), i.e. a teichoic aid. Teichoic acids were present in the walls of organisms grown under all conditions except when phosphorus limited growth. Then a different anionic polymer, composed of glucuronic acid and N-acetylgalactosamine (a teichuronic acid), was present. Under the specific growth conditions at pH7.0 and 35°C in a chemostat, teichoic acid and teichuronic acid appeared to be mutually exclusive.

scholarly journals Studies on the linkage between teichoic acid and peptidoglycan in a bacteriophage-resistant mutant of Staphylococcus aureus H

1975 ◽  
Vol 149 (3) ◽  
pp. 637-647 ◽  
Author(s):  
J E Heckels ◽  
A R Archibald ◽  
J Baddiley
Keyword(s):  
Staphylococcus Aureus ◽  
Teichoic Acid ◽  
Resistant Mutant ◽  
Muramic Acid ◽  
Glycerol Phosphate ◽  
Wall Teichoic Acid ◽  
Phosphodiester Linkage ◽  
A Chain

1. In addition to poly(ribitol phosphate) the walls of a bacteriophage-resistant mutant of Staphylococcus aureus H contain glycerol phosphate residues that are not removed on digestion with trypsin or extraction with phenol. 2. The glycerol phosphate is present in a chain, containing three or four glycerol phosphate residues, which is covalently attached to the peptidoglycan through a phosphodiester linkage to muramic acid; this linkage is readily hydrolysed by dilute alkali. 3. The degradative studies described suggest that the poly(ribitol phosphate) chains of the wall teichoic acid may be attached to the wall by linkage to this glycerol phosphate oligomer.

A protective monoclonal antibody that reacts with a novel antigen of pneumococcal teichoic acid

1987 ◽  
Vol 3 (4) ◽  
pp. 249-260 ◽  
Author(s):  
Larry S. McDaniel ◽  
W.Douglas Waltman ◽  
Barry Gray ◽  
David E. Briles
Keyword(s):  
Monoclonal Antibody ◽  
Teichoic Acid
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