Characterization of cysteine proteases from the carcinogenic liver fluke, Opisthorchis viverrini

Kaewpitoon, Natthawut, Laha, Thewarach, Kaewkes, Sasithorn, Yongvanit, Puangrat, Brindley, Paul, Loukas, Alex, and Sripa, Banchob (2008) Characterization of cysteine proteases from the carcinogenic liver fluke, Opisthorchis viverrini. Parasitology Research, 102 (4). pp. 757-764.

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Abstract

Protease activities in extracts of Opisthorchis viverrini were investigated using gelatin zymography and fluorogenic peptide substrates. Using gelatin-impregnated X-ray film, 2 microg of O. viverrini excretory-secretory products (Ov-ES) and adult somatic extract (Ov-SE) showed proteolytic activity. Zymography of both O. viverrini extracts revealed bands at approximately 30 kDa. Using fluorogenic peptide substrates, the majority of O. viverrini activity was determined to be cathepsin L-like cysteine protease (cleaved Z-Phe-Arg-aminomethylcoumarin (AMC)) whereas little or no activity was ascribable to other classes of proteases. The O. viverrini cysteine protease activity was greatest at pH 6.0 and the activity was inhibited by the class-specific inhibitors, E-64 and Z-Ala-CHN2. Chromatographic purification of O. viverrini cysteine proteases on thiol-sepharose enriched for protein(s) of approximately 30 kDa from Ov-ES and Ov-SE. The activity profile of the purified enzyme was similar to that of the cathepsin L-like activity characterized in Ov-SE and Ov-ES. Furthermore, determination of cysteine protease activity in several developmental stages of the parasite revealed the highest protease activity in metacercariae soluble extract, followed by Ov-ES, egg soluble extract, and Ov-SE. These findings demonstrated that O. viverrini has a cathepsin L-like cysteine protease(s) and suggested that abundant cysteine protease activity was present in metacercariae where the hydrolase might be involved in cyst excystation during mammalian infection.

Item ID: 36739
Item Type: Article (Research - C1)
ISSN: 1432-1955
Funders: Thailand Tropical Diseases Research Program (TTDRP) , National Institute of Health (NIH), USA, National Institute of Allergy and Infectious Diseases (NIAID), International Collaboration in Infectious Disease Research, Graduate School, Khon Kaen University, Thailand
Projects and Grants: TTDRP grant number ID02-2-HEL-05-054, International Collaboration in Infectious Disease Research award number AI065871
Date Deposited: 15 Dec 2014 00:56
FoR Codes: 11 MEDICAL AND HEALTH SCIENCES > 1108 Medical Microbiology > 110803 Medical Parasitology @ 100%
SEO Codes: 92 HEALTH > 9201 Clinical Health (Organs, Diseases and Abnormal Conditions) > 920109 Infectious Diseases @ 100%
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