Differential expression of cysteine proteases in developmental stages of the parasitic ciliate Ichthyophthirius multifiliis.

Abstract:

:An expressed sequence tag database of the freshwater fish parasite, Ichthyophthirius multifiliis (Ciliophora) was analyzed to seek for proteases potentially involved in the invasion and degradation of host tissues during infection. The translation of the database revealed two cathepsin L cysteine proteases (Icp1 and Icp2) of the C1A peptidase subfamily. The analysis of Icp1 and Icp2 sequences suggested that both proteases would be synthesized as preproproteins, with a mature domain of 27.9 and 22.8 kDa, respectively. Their expression level was determined in the trophont parasitic stage, in the tomont reproductive stage, and in the theront infective stage by real-time RT-PCR. ICP1 and ICP2 were significantly upregulated in trophont and theront stages in comparison with the tomont stage. Mature peptides of Icp1 and Icp2 were identified in crude extracts of I. multifiliis trophonts by LC-MS/MS. Zymograms showed three to seven activity bands at the optimum pH of cathepsin L cysteine proteases. Two bands displaying cysteine protease activity were identified by inhibition with E-64. They represented the major proteolytic activity of the trophont stage at pH 5-7, suggesting that cysteine proteases play an important role in the infection process.

journal_name

FEMS Microbiol Lett

authors

Jousson O,Di Bello D,Donadio E,Felicioli A,Pretti C

doi

10.1111/j.1574-6968.2006.00611.x

subject

Has Abstract

pub_date

2007-04-01 00:00:00

pages

77-84

issue

1

eissn

0378-1097

issn

1574-6968

pii

FML611

journal_volume

269

pub_type

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