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

Research output: Contribution to journalArticle

17 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)1113-1119
Number of pages7
JournalJournal of Parasitology
Volume88
Issue number6
StatePublished - Dec 2002

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Parasitology
  • Microbiology

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    Louie, K., Nordhausen, R., Robinson, T. W., Barr, B. C., & Conrad, P. A. (2002). Characterization of Neospora caninum protease, NcSUB1 (NC-p65), with rabbit anti-N54. Journal of Parasitology, 88(6), 1113-1119.