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Identification of antigenic proteins from Neospora caninum recognized by bovine immunoglobulins M, E, A and G using immunoproteomics
Author(s) -
Shin Yongseung,
Lee Eunggoo,
Shin Geewook,
Kim Youngrim,
Lee Eunyoung,
Kim Jaehoon,
Jang Hwan,
Gershwin Laurel J.,
Kim Daeyong,
Kim Yonghwan,
Kim Gonsup,
Suh Myungdeuk,
Jung Taesung
Publication year - 2004
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200400963
Subject(s) - neospora caninum , biology , microbiology and biotechnology , antigen , ribosomal protein , hsp60 , triosephosphate isomerase , antibody , chemistry , biochemistry , heat shock protein , hsp70 , enzyme , toxoplasma gondii , ribosome , rna , genetics , gene , immunology
Abstract Antigenic proteins of Neospora caninum ( N. caninum ) against bovine immunoglobulins M, E, A, and G were investigated by using immunoproteomics. Proteins of N. caninum (KBA‐2) tachyzoite lysates separated by two‐dimensional gel electrophoresis were transferred to polyvinylidene difluoride (PVDF) membranes, probed with different bovine immunoglobulin class and classified. Antigenic spots recognized were also identified by matrix‐assisted laser desorption/ionization‐time of flight‐mass spectrometry (MALDI‐TOF‐MS) analysis. 132, 84, 4, and 40 antigenic protein spots were recognized on N. caninum immunoblot profiles against bovine IgM, IgE, IgA, and IgG, respectively. Of these protein spots, the antigenic proteins recognized by either IgM, IgE, and IgG, or IgM and IgG were HSP70, pyruvate kinase, actin, NCDG‐1, tubulin α‐chain, and putative ribosomal protein S2. On the other hand, IgM, IgE, and IgA reacted with NTPase, HSP60, tubulin β‐chain, putative protein disulfide isomerase, enolase, lactate dehydrogenase, serine‐threonine phosphatase, 14‐3‐3 protein homologue, and GRA2 protein. Most of the antigenic proteins identified were associated with the process of invasion, proliferation, and egression of apicomplexans. In our study, HSP70, actin, NTPase, HSP60, pyruvate kinase, enolase, putative ribosomal protein S2, NCDG‐1, and GRA2 proteins were found to be immunodominant proteins, which may contribute to the development of diagnostic markers and vaccine.