A comparative study of the localization of wheat-germ agglutinin and its potential receptors in wheat grains

Miller, R C; Bowles, Dianna J.

doi:10.1042/bj2060571

Cited by 19 publications

(3 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In particular, the lectin wheat germ agglutinin (WGA) may play a critical role in the context of IgE receptor cross-linking. WGA is a 18 kDa lectin contained in wheat germ [38] which naturally occurs as a 36 kDa homodimer in which the two chains are linked with 16 disulfide bonds [39] . WGA is specific for N -acetyl-D-glucosamine and the chitin oligomers chitobiose and chitotriose [39] .…”

Section: Discussionmentioning

confidence: 99%

Use of Humanised Rat Basophilic Leukaemia Cell Line RS-ATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins

et al. 2014

View full text Add to dashboard Cite

BackgroundParasite-specific IgE is thought to correlate with protection against Schistosoma mansoni infection or re-infection. Only a few molecular targets of the IgE response in S. mansoni infection have been characterised. A better insight into the basic mechanisms of anti-parasite immunity could be gained from a genome-wide characterisation of such S. mansoni allergens. This would have repercussions on our understanding of allergy and the development of safe and efficacious vaccinations against helminthic parasites.Methodology/Principal FindingsA complete medium- to high-throughput amenable workflow, including important quality controls, is described, which enables the rapid translation of S. mansoni proteins using wheat germ lysate and subsequent assessment of potential allergenicity with a humanised Rat Basophilic Leukemia (RBL) reporter cell line. Cell-free translation is completed within 90 minutes, generating sufficient amounts of parasitic protein for rapid screening of allergenicity without any need for purification. Antigenic integrity is demonstrated using Western Blotting. After overnight incubation with infected individuals' serum, the RS-ATL8 reporter cell line is challenged with the complete wheat germ translation mixture and Luciferase activity measured, reporting cellular activation by the suspected allergen. The suitability of this system for characterization of novel S. mansoni allergens is demonstrated using well characterised plant and parasitic allergens such as Par j 2, SmTAL-1 and the IgE binding factor IPSE/alpha-1, expressed in wheat germ lysates and/or E. coli. SmTAL-1, but not SmTAL2 (used as a negative control), was able to activate the basophil reporter cell line.Conclusion/SignificanceThis method offers an accessible way for assessment of potential allergenicity of anti-helminthic vaccine candidates and is suitable for medium- to high-throughput studies using infected individual sera. It is also suitable for the study of the basis of allergenicity of helminthic proteins.

show abstract

Section: Discussionmentioning

confidence: 99%

Use of Humanised Rat Basophilic Leukaemia Cell Line RS-ATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins

et al. 2014

View full text Add to dashboard Cite

show abstract

“…It is a dimeric protein with four binding sites that has specificity for GlcNAc (5) and its -(1 4) linked oligosaccharides and analogous sugars [113]. It has also been shown to have affinity for sialic acid (neuraminic acid) (9) [111], which is present in a wide variety of glycans as an end cap bonded to Gal (1) or GalNAc (6) [105].…”

Section: Lectinsmentioning

confidence: 99%

Mass Spectrometry in the Elucidation of the Glycoproteome of Bacterial Pathogens

Graham

Hess

2010

View full text Add to dashboard Cite

Presently some three hundred post-translational modifications are known to occur in bacteria in vivo. Many of these modifications play critical roles in the regulation of proteins and control key biological processes. One of the most predominant modifications, N-and O-glycosylations are now known to be present in bacteria (and archaea) although they were long believed to be limited to eukaryotes. In a number of human pathogens these glycans have been found attached to the surfaces of pilin, flagellin and other surface and secreted proteins where it has been demonstrated that they play a role in the virulence of these bacteria. Mass spectrometry characterization of these glycosylation events has been the enabling key technology for these findings. This review will look at the use of mass spectrometry as a key technology for the detection and mapping of these modifications within microorganisms, with particular reference to the human pathogens, Campylobacter jejuni and Mycobacterium tuberculosis. The overall aim of this review will be to give a basic understanding of the current 'state-of-the-art' of the key techniques, principles and technologies, including bioinformatics tools, involved in the analysis of the glycosylation modifications.

show abstract

“…However, effectiveness is dependent on the relative abundance of the binding component and some idea as to an appropriate separation system, especially if the components are not glycoproteins or glycolipids. Similarily, immobilized lectin systems have been used with great success for characterization of glycoproteins from a wide variety of cell types, but their use in identification of potential 'receptors' within systems from which the lectin was isolated has been limited (Gansera et al, 1979;Breuer & Siu, 1981; Bowles & Marcus, 1981;Ray & Lerner, 1982;Miller & Bowles, 1982;Marcus et al, 1984).…”

mentioning

confidence: 99%

Interactions of soya-bean agglutinin with purified glycoconjugates and soya-bean seed components

Bond¹,

Chaplin²,

Bowles³

1985

Biochemical Journal

Self Cite

View full text Add to dashboard Cite

A radioaffinity assay for lectin binding to receptors was developed and characterized by using the interactions between soya-bean agglutinin and four glycoconjugates, namely thyroglobulin, galactomannan, fetuin and asialofetuin. On application of the assay to soya-bean extracts a wide range of seed components were found to have the capacity to interact with soya-bean agglutinin. These included both trichloroacetic acid-soluble and trichloroacetic acid-insoluble glycoconjugates and two classes of particulate matter distinguished by their differential solubility in Triton X-100.

show abstract

A comparative study of the localization of wheat-germ agglutinin and its potential receptors in wheat grains

Cited by 19 publications

References 30 publications

Use of Humanised Rat Basophilic Leukaemia Cell Line RS-ATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins

Use of Humanised Rat Basophilic Leukaemia Cell Line RS-ATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins

Mass Spectrometry in the Elucidation of the Glycoproteome of Bacterial Pathogens

Interactions of soya-bean agglutinin with purified glycoconjugates and soya-bean seed components

Contact Info

Product

Resources

About