CHARACTERIZATION OF NEOSPORA CANINUM PROTEASE, NcSUB1 (NC-p65), WITH RABBIT ANTI-N54

Kitland Louie; Robert Nordhausen; Thomas W. Robinson; Bradd C. Barr; Patricia A. Conrad

doi:10.1645/0022-3395(2002)088[1113:CONCPN]2.0.CO;2

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1 December 2002 CHARACTERIZATION OF NEOSPORA CANINUM PROTEASE, NcSUB1 (NC-p65), WITH RABBIT ANTI-N54

Kitland Louie, Robert Nordhausen, Thomas W. Robinson, Bradd C. Barr, Patricia A. Conrad

Author Affiliations +

J. of Parasitology, 88(6):1113-1119 (2002). https://doi.org/10.1645/0022-3395(2002)088[1113:CONCPN]2.0.CO;2

Abstract

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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Kitland Louie, Robert Nordhausen, Thomas W. Robinson, Bradd C. Barr and Patricia A. Conrad "CHARACTERIZATION OF NEOSPORA CANINUM PROTEASE, NcSUB1 (NC-p65), WITH RABBIT ANTI-N54," Journal of Parasitology 88(6), (1 December 2002). https://doi.org/10.1645/0022-3395(2002)088[1113:CONCPN]2.0.CO;2

Received: 8 January 2002; Accepted: 1 June 2002; Published: 1 December 2002

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