title: Glycoproteomic studies of IgE from a novel hyper IgE syndrome linked to PGM3 mutation.
abstract: Glycans serve as important regulators of antibody activities and half-lives. IgE is the most heavily glycosylated antibody, but in comparison to other antibodies little is known about its glycan structure function relationships. We therefore describe the site specific IgE glycosylation from a patient with a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation. A two-step method was developed to prepare two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis as a control subject. Then, a glycoproteomic strategy was used to study the site-specific glycosylation. No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. The results showed similar glycosylation profiles between the two IgE samples. These observations, together with previous knowledge of IgE glycosylation, imply that IgE glycosylation is similarly regulated among healthy control, allergy and PGM3 related hyper IgE syndrome.
authors.affiliation: Department of Life Sciences, Imperial College London, South Kensington Campus, London, SW7 2AZ, UK.|Division of Cell Signalling and Immunology, School of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK.
authors.forename: Gang
authors.initials: G
authors.lastname: Wu
authors.affiliation: Department of Life Sciences, Imperial College London, South Kensington Campus, London, SW7 2AZ, UK.
authors.forename: Paul G
authors.initials: PG
authors.lastname: Hitchen
authors.affiliation: Department of Life Sciences, Imperial College London, South Kensington Campus, London, SW7 2AZ, UK.
authors.forename: Maria
authors.initials: M
authors.lastname: Panico
authors.affiliation: Department of Life Sciences, Imperial College London, South Kensington Campus, London, SW7 2AZ, UK.
authors.forename: Simon J
authors.initials: SJ
authors.lastname: North
authors.affiliation: Laboratory of Immunopathology, Vaccinology and Molecular Genetics, Pasteur Institute of Tunis and University Tunis El Manar, Tunis, Tunisia.
authors.forename: Mohamed-Ridha
authors.initials: MR
authors.lastname: Barbouche
authors.affiliation: MS-RTC (Mass Spectrometry Research and Training Centre), Suite 3.1 Lido Medical Centre, St. Saviours Road, Jersey, JE2 7LA, UK.
authors.forename: Daniel
authors.initials: D
authors.lastname: Binet
authors.affiliation: Department of Life Sciences, Imperial College London, South Kensington Campus, London, SW7 2AZ, UK.|MS-RTC (Mass Spectrometry Research and Training Centre), Suite 3.1 Lido Medical Centre, St. Saviours Road, Jersey, JE2 7LA, UK.
authors.forename: Howard R
authors.initials: HR
authors.lastname: Morris
authors.affiliation: Department of Life Sciences, Imperial College London, South Kensington Campus, London, SW7 2AZ, UK.
authors.forename: Anne
authors.initials: A
authors.lastname: Dell
authors.affiliation: Department of Life Sciences, Imperial College London, South Kensington Campus, London, SW7 2AZ, UK. s.haslam@imperial.ac.uk.|Department of Life Sciences, Faculty of Natural Sciences, Imperial College London, South Kensington Campus, London, SW7 2AZ, UK. s.haslam@imperial.ac.uk.
authors.forename: Stuart M
authors.initials: SM
authors.lastname: Haslam
mesh_terms: D001665:Binding Sites; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007073:Immunoglobulin E / Q000737:chemistry / Q000378:metabolism*; D007589:Job Syndrome / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods; D025202:Molecular Diagnostic Techniques / Q000379:methods; D009154:Mutation; D010733:Phosphoglucomutase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000737:chemistry / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D020543:Proteome; D007073:Immunoglobulin E; C549916:PGM3 protein, human; D010733:Phosphoglucomutase
keywords: Glycoproteomics; Hyper IgE Syndrome; IgE; Phosphoglucomutase 3