The cathepsin B of Toxoplasma gondii, toxopain-1, is critical for parasite invasion and rhoptry protein processing

Xuchu Que, Huân Ngô, Jeffrey Lawton, Mary Gray, Qing Liu, Juan Engel, Linda Brinen, Partho Ghosh, Keith A Joiner, Sharon L. Reed

Research output: Contribution to journalArticle

77 Citations (Scopus)

Abstract

Cysteine proteinases play a major role in invasion and intracellular survival of a number of pathogenic parasites. We cloned a single copy gene, tgcpl, from Toxoplasma gondii and refolded recombinant enzyme to yield active proteinase. Substrate specificity of the enzyme and homology modeling identified the proteinase as a cathepsin B. Specific cysteine proteinase inhibitors interrupted invasion by tachyzoites. The T. gondii cathepsin B localized to rhoptries, secretory organelles required for parasite invasion into cells. Processing of the pro-rhoptry protein 2 to mature rhoptry proteins was delayed by incubation of extracellular parasites with a cathepsin B inhibitor prior to pulse-chase immunoprecipitation. Delivery of cathepsin B to mature rhoptries was impaired in organisms with disruptions in rhoptry formation by expression of a dominant negative μ1-adaptin. Similar disruption of rhoptry formation was observed when infected fibroblasts were treated with a specific inhibitor of cathepsin B, generating small and poorly developed rhoptries. This first evidence for localization of a cysteine proteinase to the unusual rhoptry secretory organelle of an apicomplexan parasite suggests that the rhoptries may be a prototype of a lysosome-related organelle and provides a critical link between cysteine proteinases and parasite invasion for this class of organism.

Original languageEnglish (US)
Pages (from-to)25791-25797
Number of pages7
JournalJournal of Biological Chemistry
Volume277
Issue number28
DOIs
StatePublished - Jul 12 2002
Externally publishedYes

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Cathepsin B
Parasites
Cysteine Proteases
Organelles
Processing
Toxoplasma
Proteins
Peptide Hydrolases
Cysteine Proteinase Inhibitors
Enzymes
Fibroblasts
Substrate Specificity
Lysosomes
Immunoprecipitation
Genes
Toxoplasma gondii toxopain-1
Substrates

ASJC Scopus subject areas

  • Biochemistry

Cite this

The cathepsin B of Toxoplasma gondii, toxopain-1, is critical for parasite invasion and rhoptry protein processing. / Que, Xuchu; Ngô, Huân; Lawton, Jeffrey; Gray, Mary; Liu, Qing; Engel, Juan; Brinen, Linda; Ghosh, Partho; Joiner, Keith A; Reed, Sharon L.

In: Journal of Biological Chemistry, Vol. 277, No. 28, 12.07.2002, p. 25791-25797.

Research output: Contribution to journalArticle

Que, X, Ngô, H, Lawton, J, Gray, M, Liu, Q, Engel, J, Brinen, L, Ghosh, P, Joiner, KA & Reed, SL 2002, 'The cathepsin B of Toxoplasma gondii, toxopain-1, is critical for parasite invasion and rhoptry protein processing', Journal of Biological Chemistry, vol. 277, no. 28, pp. 25791-25797. https://doi.org/10.1074/jbc.M202659200
Que, Xuchu ; Ngô, Huân ; Lawton, Jeffrey ; Gray, Mary ; Liu, Qing ; Engel, Juan ; Brinen, Linda ; Ghosh, Partho ; Joiner, Keith A ; Reed, Sharon L. / The cathepsin B of Toxoplasma gondii, toxopain-1, is critical for parasite invasion and rhoptry protein processing. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 28. pp. 25791-25797.
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