Comparison of proteome and antigenic proteome between two Neospora caninum isolates.
This study was conducted to explore the relationship between two isolates of Neospora caninum (N. caninum) (KBA-2 and VMDL-1) using proteomics. To achieve the goal, proteins of N. caninum tachyzoite lysates of KBA-2 and VMDL-1 were separated by two-dimensional gel electrophoresis (2-DE), stained with silver-nitrate and analyzed using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) to compare protein profiles. In addition, proteins separated by 2-DE were transferred to membranes, probed with bovine anti-N. caninum KBA-2 immunoglobulin G, and reactive proteins were visualized and compared between the two isolates. Most spots on 2-DE profiles and antigenic spots on 2-DE immunoblot profiles were located at similar locations in terms of isoelectric point and molecular weight. Proteins common to both isolates included the following: heat shock protein 70, subtilisin-like serine protease, nucleoside triphosphatase, heat shock protein 60, pyruvate kinase, tubulin alpha, tubulin beta, enolase, putative protein disulfide isomerase, actin, fructase-1,6-bisphosphatase, putative ribosomal protein S2, microneme protein Nc-P38, lactate dihydrogenase, fructose-1,6-bisphosphatase aldolase, serine threonine phosphatase 2C, 14-3-3 protein homologue, N. caninum dense granule-1 and NcGRA2. As a consequence, even though N. caninum KBA-2 and VMDL-1 isolates were isolated from geographically distinct locations there were significant homology in the proteome and antigenic proteome profiles. In addition, proteomic approach was verified as a useful tool for understanding of host immune response against different isolates of protozoa.
Shin
Yong-Seung
YS
Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju, Gyeong-nam 660-701, Republic of Korea.
Shin
Gee-Wook
GW
Kim
Young-Rim
YR
Lee
Eun-Young
EY
Yang
Hyang-Hee
HH
Palaksha
K J
KJ
Youn
Hee-Jeong
HJ
Kim
Jae-Hoon
JH
Kim
Dae-Yong
DY
Marsh
A E
AE
Lakritz
J
J
Jung
Tae-Sung
TS
D000818:Animals; D000953:Antigens, Protozoan / Q000032:analysis / Q000378:metabolism*; D002417:Cattle; D015180:Electrophoresis, Gel, Two-Dimensional / Q000662:veterinary; D005260:Female; D015151:Immunoblotting / Q000662:veterinary; D007526:Isoelectric Point; D008970:Molecular Weight; D018688:Neospora / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D040901:Proteomics; D015800:Protozoan Proteins / Q000032:analysis / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000662:veterinary
D000953:Antigens, Protozoan; D015800:Protozoan Proteins