Identification, recombinant protein production, and functional analysis of a M60-like metallopeptidase, secreted by the liver fluke Opisthorchis viverrini

Ta, Binh T.T., Nguyen, D. Linh, Jala, Isabelle, Dontumprai, Rieofarng, Plumworasawat, Sirikanya, Aighewi, Omorose, Ong, Emily, Shawley, Audrey, Potriquet, Jeremy, Saichua, Prasert, van Diepen, Angela, Sripa, Banchob, Hokke, Cornelis H., and Suttiprapa, Sutas (2020) Identification, recombinant protein production, and functional analysis of a M60-like metallopeptidase, secreted by the liver fluke Opisthorchis viverrini. Parasitology International, 75. 102050.

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Abstract

The carcinogenic liver fluke Opisthorchis viverrini (O. viverrini) is endemic in Thailand and neighboring countries including Laos PDR, Vietnam and Cambodia. Infections with O. viverrini lead to hepatobiliary abnormalities including bile duct cancer-cholangiocarcinoma (CCA). Despite decades of extensive studies, the underlying mechanisms of how this parasite survives in the bile duct and causes disease are still unclear. Therefore, this study aims to identify and characterize the most abundant protein secreted by the parasite. Proteomics and bioinformatics analysis revealed that the most abundant secretory protein is a metallopeptidase, named Ov-M60-like-1. This protein contains an N-terminal carbohydrate-binding domain and a C-terminal M60-like domain with a zinc metallopeptidase HEXXH motif. Further analysis by mass spectrometry revealed that Ov-M60-like-1 is N-glycosylated. Recombinant Ov-M60-like-1 (rOv-M60-like-1) expressed in Escherichia coli (E. coli) was able to digest bovine submaxillary mucin (BSM). The mucinase activity was inhibited by the ion chelating agent EDTA, confirming its metallopeptidase identity. The enzyme was active at temperatures ranging 25-37 degrees C in a broad pH range (pH 2-10). The identification of Ov-M60-like-1 mucinase as the major secretory protein of O. viverrini worms warrants further research into the role of this glycoprotein in the pathology induced by this carcinogenic worm.

Item ID: 62677
Item Type: Article (Research - C1)
ISSN: 1873-0329
Keywords: Opisthorchis viverrini, Liver fluke, M60-like metallopeptidase, Mucinase, Glycoprotein
Copyright Information: © 2020 Elsevier B.V. All rights reserved.
Funders: Khon Kaen University (KKU), Thailand Research Fund (TRF), Mahidol University, Mekong Health Science Research Institute
Projects and Grants: GMS-KKU Scholarship, TRF TRG5780147
Date Deposited: 01 Apr 2020 07:40
FoR Codes: 32 BIOMEDICAL AND CLINICAL SCIENCES > 3207 Medical microbiology > 320704 Medical parasitology @ 30%
31 BIOLOGICAL SCIENCES > 3104 Evolutionary biology > 310407 Host-parasite interactions @ 70%
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