Establishment of a two-dimensional electrophoresis map for Neospora caninum tachyzoites by proteomics.
Expressed proteins and antigens from Neospora caninum tachyzoites were studied by two-dimensional gel electrophoresis and immunoblot analysis combined with matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Thirty-one spots corresponding to 20 different proteins were identified from N. caninum tachyzoites by peptide mass fingerprinting. Six proteins were identified from a N. caninum database (NTPase, 14-3-3 protein homologue, NcMIC1, NCDG1, NcGRA1 and NcGRA2), and 11 proteins were identified in closely related species using the T. gondii database (HSP70, HSP60, pyruvate kinase, tubulin alpha- and beta-chain, putative protein disulfide isomerase, enolase, actin, fructose-1,6-bisphosphatase, lactate dehydrogenase and glyceradehyde-3-phosphate dehydrogenase). One hundred and two antigen spots were observed using pH 4-7 IPG strips on immunoblot profiles. Among them, 17 spots corresponding to 11 antigenic proteins were identified from a N. caninum protein map. This study involved the construction of in-depth protein maps for N. caninum tachyzoites, which will be of value for studies of its pathogenesis, drug and vaccine development, and phylogenetic studies.
Lee
Eung Goo
EG
Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, 900 Gazwa-Dong, Jinju, Gyeong-nam 660-701, South Korea.
Kim
Jae Hoon
JH
Shin
Yong Seung
YS
Shin
Gee Wook
GW
Suh
Myung Deuk
MD
Kim
Dae Yong
DY
Kim
Yong Hwan
YH
Kim
Gon Sup
GS
Jung
Tae Sung
TS
D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional; D007525:Isoelectric Focusing; D018688:Neospora / Q000737:chemistry* / Q000276:immunology; D020543:Proteome / Q000737:chemistry*; D015800:Protozoan Proteins / Q000737:chemistry* / Q000276:immunology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000913:Antibodies, Protozoan; D000954:Antigens, Surface; D020543:Proteome; D015800:Protozoan Proteins