TY - JOUR
T1 - Analysis of the glycoproteome of Toxoplasma gondii using lectin affinity chromatography and tandem mass spectrometry
AU - Luo, Qilie
AU - Upadhya, Rajendra
AU - Zhang, Hong
AU - Madrid-Aliste, Carlos
AU - Nieves, Edward
AU - Kim, Kami
AU - Angeletti, Ruth Hogue
AU - Weiss, Louis M.
N1 - Funding Information:
Supported by NIH/NIAID grant AI31744 (LMW), NIH/NIAID contract HHSN266200400054C, NIH/NIAID R01AI087625 (KK) and RC4AI092801 (KK).
PY - 2011/12
Y1 - 2011/12
N2 - 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.
AB - 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.
KW - Glycoproteome
KW - Glycosylation
KW - Lectin chromatography
KW - Membrane proteins
KW - Toxoplasma
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U2 - 10.1016/j.micinf.2011.08.013
DO - 10.1016/j.micinf.2011.08.013
M3 - Article
C2 - 21920448
AN - SCOPUS:81155132158
SN - 1286-4579
VL - 13
SP - 1199
EP - 1210
JO - Microbes and Infection
JF - Microbes and Infection
IS - 14-15
ER -