title: Analysis of the glycoproteome of Toxoplasma gondii using lectin affinity chromatography and tandem mass spectrometry.
abstract: Glycoproteins are involved in many important molecular recognition processes including invasion, adhesion, differentiation, and development. To identify the glycoproteins of Toxoplasma gondii, a proteomic analysis was undertaken. T. gondii proteins were prepared and fractioned using lectin affinity chromatography. The proteins in each fraction were then separated using SDS-PAGE and identified by tryptic in gel digestion followed by tandem mass spectrometry. Utilizing these methods 132 proteins were identified. Among the identified proteins were 17 surface proteins, 9 microneme proteins, 15 rhoptry proteins, 11 heat shock proteins (HSP), and 32 hypothetical proteins. Several proteins had 1-5 transmembrane domains (TMD) with some being as large as 608.3 kDa. Both lectin-fluorescence labeling and lectin blotting were employed to confirm the presence of carbohydrates on the surface or cytoplasm of T. gondii parasites. PCR demonstrated that selected hypothetical proteins were expressed in T. gondii tachyzoites. This data provides a large-scale analysis of the T. gondii glycoproteome. Studies of the function of glycosylation of these proteins may help elucidate mechanism(s) involved in invasion improving drug therapy as well as identify glycoproteins that may prove to be useful as vaccine candidates.
authors.affiliation: Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: Qilie
authors.initials: Q
authors.lastname: Luo
authors.forename: Rajendra
authors.initials: R
authors.lastname: Upadhya
authors.forename: Hong
authors.initials: H
authors.lastname: Zhang
authors.forename: Carlos
authors.initials: C
authors.lastname: Madrid-Aliste
authors.forename: Edward
authors.initials: E
authors.lastname: Nieves
authors.forename: Kami
authors.initials: K
authors.lastname: Kim
authors.forename: Ruth Hogue
authors.initials: RH
authors.lastname: Angeletti
authors.forename: Louis M
authors.initials: LM
authors.lastname: Weiss
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D015153:Blotting, Western; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000469:parasitology; D054794:Glycomics; D006023:Glycoproteins / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D037102:Lectins / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D040901:Proteomics; D053719:Tandem Mass Spectrometry; D014122:Toxoplasma / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D014123:Toxoplasmosis / Q000469:parasitology; D014357:Trypsin / Q000378:metabolism
chemical_list: D000953:Antigens, Protozoan; D006023:Glycoproteins; D037102:Lectins; D014357:Trypsin