The functional expression and characterisation of a cysteine peptidase from the invasive stage of the neuropathogenic schistosome Trichobilharzia regenti.

International Journal for Parasitology
Katerina DoleckováP Horák

Abstract

A transcriptional product of a gene encoding cathepsin B-like peptidase in the bird schistosome Trichobilharzia regenti was identified and cloned. The enzyme was named TrCB2 due to its 77% sequence similarity to cathepsin B2 from the important human parasite Schistosoma mansoni. The zymogen was expressed in the methylotropic yeast Pichia pastoris; procathepsin B2 underwent self-processing in yeast media. The peptidolytic activity of the recombinant enzyme was characterised using synthetic fluorogenic peptide substrates at optimal pH 6.0. Functional studies using different specific inhibitors proved the typical cathepsin B-like nature of the enzyme. The S(2) subsite specificity profile of recombinant TrCB2 was obtained. Using monospecific antibodies against the recombinant enzyme, the presence of cathepsin B2 was confirmed in extracts from cercariae (infective stage) and schistosomula (early post-cercarial stage) of T. regenti on Western blots. Also, cross-reactivity was observed between T. regenti and S. mansoni cathepsins B2 in extracts of cercariae, schistosomula or adults. In T. regenti, the antisera localised the enzyme to post-acetabular penetration glands of cercariae implying an important role in the penetration of host ...Continue Reading

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Citations

Feb 24, 2011·Parasitology Research·Yan-ping HanZhi-yue Lv
Aug 24, 2012·Clinical Reviews in Allergy & Immunology·Libuše KolářováMichael Doenhoff
Apr 18, 2012·PLoS Neglected Tropical Diseases·Jessica R IngramJames H McKerrow
Jun 29, 2010·Trends in Parasitology·Peter M SmookerTerry W Spithill
Nov 6, 2012·Journal of Parasitology Research·Lucie LichtenbergováHorák Petr
Aug 24, 2018·PLoS Neglected Tropical Diseases·Alexandra GroteSara Lustigman
Jan 9, 2015·Clinical Microbiology Reviews·Petr HorákSara Vanessa Brant
Mar 8, 2020·Parasite Immunology·Martin MajerPetr Horák
Oct 27, 2020·PLoS Neglected Tropical Diseases·Bingkuan ZhuWei Hu

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Datasets Mentioned

BETA
AY226984

Methods Mentioned

BETA
PCRs
PCR
electrophoresis
glycosylation
confocal microscopy

Software Mentioned

BLASTp
ClustalW
SignalP
ExPASy

Related Concepts