NcSUB1 (formerly known as NC-p65) is the first molecularly described proteolytic enzyme of the intracellular protozoan parasite Neospora caninum. This report describes the characterization of a rabbit anti-N54, which is an antiserum generated against an internal fragment of NcSUB1 (amino acids 649–783). In immunofluorescence studies rabbit anti-N54 labeled the apical end of the fixed parasite. By immuno-gold electron microscopy, the antibody bound primarily to the microneme organelles of the parasite. Analysis of secreted parasitic proteins indicated that a protein of molecular weight 65 kDa (reduced) or 55 kDa (nonreduced) was recognized by the antibody. The same secreted proteins were affinity purified with rabbit anti-N54–coupled resins and were shown to contain major proteolytic activity by zymography. Thus, rabbit anti-N54 is the first antibody developed for N. caninum that binds to the microneme proteins and recognizes a major secreted enzyme.
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December 2002
Research Article|
December 01 2002
CHARACTERIZATION OF NEOSPORA CANINUM PROTEASE, NcSUB1 (NC-p65), WITH RABBIT ANTI-N54
Kitland Louie;
Kitland Louie
Department of Pathology, Microbiology, and Immunology, School of Veterinary Medicine, University of California, One Shields Avenue, Davis, California 95616. paconrad@ucdavis.edu
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Robert Nordhausen;
Robert Nordhausen
Department of Pathology, Microbiology, and Immunology, School of Veterinary Medicine, University of California, One Shields Avenue, Davis, California 95616. paconrad@ucdavis.edu
* California Animal Health and Food Safety Laboratory, School of Veterinary Medicine, University of California, Davis, California 95616
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Thomas W. Robinson;
Thomas W. Robinson
Department of Pathology, Microbiology, and Immunology, School of Veterinary Medicine, University of California, One Shields Avenue, Davis, California 95616. paconrad@ucdavis.edu
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Bradd C. Barr;
Bradd C. Barr
Department of Pathology, Microbiology, and Immunology, School of Veterinary Medicine, University of California, One Shields Avenue, Davis, California 95616. paconrad@ucdavis.edu
* California Animal Health and Food Safety Laboratory, School of Veterinary Medicine, University of California, Davis, California 95616
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Patricia A. Conrad
Patricia A. Conrad
Department of Pathology, Microbiology, and Immunology, School of Veterinary Medicine, University of California, One Shields Avenue, Davis, California 95616. paconrad@ucdavis.edu
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J Parasitol (2002) 88 (6): 1113–1119.
Citation
Kitland Louie, Robert Nordhausen, Thomas W. Robinson, Bradd C. Barr, Patricia A. Conrad; CHARACTERIZATION OF NEOSPORA CANINUM PROTEASE, NcSUB1 (NC-p65), WITH RABBIT ANTI-N54. J Parasitol 1 December 2002; 88 (6): 1113–1119. doi: https://doi.org/10.1645/0022-3395(2002)088[1113:CONCPN]2.0.CO;2
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