Analysis of the glycoproteome of Toxoplasma gondii using lectin affinity chromatography and tandem mass spectrometry

Qilie Luo, Rajendra Upadhya, Hong Zhang, Carlos Madrid-Aliste, Edward Nieves, Kami Kim, Ruth Hogue Angeletti, Louis M. Weiss

Research output: Contribution to journalArticle

25 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)1199-1210
Number of pages12
JournalMicrobes and Infection
Volume13
Issue number14-15
DOIs
StatePublished - Dec 2011

Fingerprint

Toxoplasma
Tandem Mass Spectrometry
Affinity Chromatography
Lectins
Proteins
Glycoproteins
Heat-Shock Proteins
Glycosylation
Proteomics
Polyacrylamide Gel Electrophoresis
Digestion
Membrane Proteins
Cytoplasm
Parasites
Vaccines
Fluorescence
Gels
Carbohydrates
Drug Therapy
Polymerase Chain Reaction

Keywords

  • Glycoproteome
  • Glycosylation
  • Lectin chromatography
  • Membrane proteins
  • Toxoplasma

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Infectious Diseases

Cite this

Analysis of the glycoproteome of Toxoplasma gondii using lectin affinity chromatography and tandem mass spectrometry. / Luo, Qilie; Upadhya, Rajendra; Zhang, Hong; Madrid-Aliste, Carlos; Nieves, Edward; Kim, Kami; Angeletti, Ruth Hogue; Weiss, Louis M.

In: Microbes and Infection, Vol. 13, No. 14-15, 12.2011, p. 1199-1210.

Research output: Contribution to journalArticle

Luo, Qilie ; Upadhya, Rajendra ; Zhang, Hong ; Madrid-Aliste, Carlos ; Nieves, Edward ; Kim, Kami ; Angeletti, Ruth Hogue ; Weiss, Louis M. / Analysis of the glycoproteome of Toxoplasma gondii using lectin affinity chromatography and tandem mass spectrometry. In: Microbes and Infection. 2011 ; Vol. 13, No. 14-15. pp. 1199-1210.
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