Identification, recombinant protein production, and functional analysis of a M60-like metallopeptidase, secreted by the liver fluke Opisthorchis viverrini
| Autor: | Jeremy Potriquet, Binh T.T. Ta, Cornelis H. Hokke, Prasert Saichua, Sutas Suttiprapa, Isabelle Jala, Rieofarng Dontumprai, Sirikanya Plumworasawat, Angela van Diepen, Banchob Sripa, D. Linh Nguyen, Emily Ong, Omorose Aighewi, Audrey Shawley |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Metallopeptidase 030231 tropical medicine Proteomics medicine.disease_cause Opisthorchiasis Microbiology 03 medical and health sciences 0302 clinical medicine parasitic diseases medicine Animals Opisthorchis viverrini M60-like metallopeptidase Amino Acid Sequence Escherichia coli Phylogeny chemistry.chemical_classification biology Opisthorchis Mucin Liver fluke Helminth Proteins 030108 mycology & parasitology biology.organism_classification Recombinant Proteins Infectious Diseases Secretory protein chemistry Mucinase Metalloproteases Parasitology Glycoprotein Sequence Alignment |
| Zdroj: | Parasitology International, 75. ELSEVIER IRELAND LTD |
| ISSN: | 1873-0329 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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