title: The PCome of Ascaris suum as a model system for intestinal nematodes: identification of phosphorylcholine-substituted proteins and first characterization of the PC-epitope structures.
abstract: In multicellular parasites (e.g., nematodes and protozoa), proteins and glycolipids have been found to be decorated with phosphorylcholine (PC). PC can provoke various effects on immune cells leading to an immunomodulation of the host's immune system. This immunomodulation allows long-term persistence but also prevents severe pathology due to downregulation of cellular immune responses. PC-containing antigens have been found to interfere with key proliferative signaling pathways in B and T cells, development of dendritic cells and macrophages, and mast cell degranulation. These effects contribute to the observed modulated cytokine levels and impairment of lymphocyte proliferation. In contrast to glycosphingolipids, little is known about the PC-epitopes of proteins. So far, only a limited number of PC-modified proteins from nematodes have been identified. In this project, PC-substituted proteins and glycolipids in Ascaris suum have been localized by immunohistochemistry in specific tissues of the body wall, intestine, and reproductive tract. Subsequently, we investigated the PCome of A. suum by 2D gel-based proteomics and detection by Western blotting using the PC-specific antibody TEPC-15. By peptide-mass-fingerprint matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), we could identify 59 PC-substituted proteins, which are in involved multiple cellular processes. In addition to membrane proteins like vitellogenin-6, we found proteins with structural (e.g., tubulins) and metabolic (e.g., pyruvate dehydrogenase) functions or which can act in the defense against the host's immune response (e.g., serpins). Initial characterization of the PC-epitopes revealed a predominant linkage of PC to the proteins via N-glycans. Our data form the basis for more detailed investigations of the PC-epitope structures as a prerequisite for comprehensive understanding of the molecular mechanisms of immunomodulation.
authors.affiliation: Protein Analytics, Institute of Biochemistry, Faculty of Medicine, Justus-Liebig University, Friedrichstrasse 24, 35392, Giessen, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Timm
authors.affiliation: Protein Analytics, Institute of Biochemistry, Faculty of Medicine, Justus-Liebig University, Friedrichstrasse 24, 35392, Giessen, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Grabitzki
authors.affiliation: Protein Analytics, Institute of Biochemistry, Faculty of Medicine, Justus-Liebig University, Friedrichstrasse 24, 35392, Giessen, Germany.
authors.forename: Cinar
authors.initials: C
authors.lastname: Severcan
authors.affiliation: Protein Analytics, Institute of Biochemistry, Faculty of Medicine, Justus-Liebig University, Friedrichstrasse 24, 35392, Giessen, Germany.
authors.forename: Suzan
authors.initials: S
authors.lastname: Muratoglu
authors.affiliation: Protein Analytics, Institute of Biochemistry, Faculty of Medicine, Justus-Liebig University, Friedrichstrasse 24, 35392, Giessen, Germany.
authors.forename: Lisa
authors.initials: L
authors.lastname: Ewald
authors.affiliation: Protein Analytics, Institute of Biochemistry, Faculty of Medicine, Justus-Liebig University, Friedrichstrasse 24, 35392, Giessen, Germany.
authors.forename: Yavuz
authors.initials: Y
authors.lastname: Yilmaz
authors.affiliation: Protein Analytics, Institute of Biochemistry, Faculty of Medicine, Justus-Liebig University, Friedrichstrasse 24, 35392, Giessen, Germany. guenter.lochnit@biochemie.med.uni-giessen.de.
authors.forename: Guenter
authors.initials: G
authors.lastname: Lochnit
mesh_terms: D000818:Animals; D000947:Antigens, Helminth / Q000737:chemistry* / Q000276:immunology; D017165:Ascaris suum / Q000737:chemistry* / Q000276:immunology; D015153:Blotting, Western; D015180:Electrophoresis, Gel, Two-Dimensional; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology; D005260:Female; D015801:Helminth Proteins / Q000737:chemistry* / Q000276:immunology; D056747:Immunomodulation; D008954:Models, Biological; D010767:Phosphorylcholine / Q000737:chemistry* / Q000276:immunology; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000947:Antigens, Helminth; D000939:Epitopes; D015801:Helminth Proteins; D010767:Phosphorylcholine
keywords: Immunomodulation; MALDI-TOF-MS; Nematodes; Phosphorylcholine; Proteomics