Cathepsin F cysteine protease of the human liver fluke, Opisthorchis viverrini
The liver fluke Opisthorchis viverrini is classified as a class I carcinogen due to the association between cholangiocarcinoma and chronic O. viverrini infection. During its feeding activity within the bile duct, the parasite secretes several cathepsin F cysteine proteases that may induce or contrib...
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| Veröffentlicht in: | PLoS neglected tropical diseases 2009, Vol.3 (3), p.e398 |
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| creator | Pinlaor, Porntip Kaewpitoon, Natthawut Laha, Thewarach Sripa, Banchob Kaewkes, Sasithorn Morales, Maria E Mann, Victoria H Parriott, Sandi K Suttiprapa, Sutas Robinson, Mark W To, Joyce Dalton, John P Loukas, Alex Brindley, Paul J |
| description | The liver fluke Opisthorchis viverrini is classified as a class I carcinogen due to the association between cholangiocarcinoma and chronic O. viverrini infection. During its feeding activity within the bile duct, the parasite secretes several cathepsin F cysteine proteases that may induce or contribute to the pathologies associated with hepatobiliary abnormalities.
Here, we describe the cDNA, gene organization, phylogenetic relationships, immunolocalization, and functional characterization of the cathepsin F cysteine protease gene, here termed Ov-cf-1, from O. viverrini. The full length mRNA of 1020 nucleotides (nt) encoded a 326 amino acid zymogen consisting of a predicted signal peptide (18 amino acids, aa), prosegment (95 aa), and mature protease (213 aa). BLAST analysis using the Ov-CF-1 protein as the query revealed that the protease shared identity with cathepsin F-like cysteine proteases of other trematodes, including Clonorchis sinensis (81%), Paragonimus westermani (58%), Schistosoma mansoni and S. japonicum (52%), and with vertebrate cathepsin F (51%). Transcripts encoding the protease were detected in all developmental stages that parasitize the mammalian host. The Ov-cf-1 gene, of approximately 3 kb in length, included seven exons interrupted by six introns; the exons ranged from 69 to 267 bp in length, the introns from 43 to 1,060 bp. The six intron/exon boundaries of Ov-cf-1 were conserved with intron/exon boundaries in the human cathepsin F gene, although the gene structure of human cathepsin F is more complex. Unlike Ov-CF-1, human cathepsin F zymogen includes a cystatin domain in the prosegment region. Phylogenetic analysis revealed that the fluke, human, and other cathepsin Fs branched together in a clade discrete from the cathepsin L cysteine proteases. A recombinant Ov-CF-1 zymogen that displayed low-level activity was expressed in the yeast Pichia pastoris. Although the recombinant protease did not autocatalytically process and activate to a mature enzyme, trans-processing by Fasciola hepatica cathepsin L cleaved the prosegment of Ov-CF-1, releasing a mature cathepsin F with activity against the peptide Z-Phe-Arg-NHMec >50 times that of the zymogen. Immunocytochemistry using antibodies raised against the recombinant enzyme showed that Ov-CF-1 is expressed in the gut of the mature hermaphroditic fluke and also in the reproductive structures, including vitelline glands, egg, and testis. Ov-CF-1 was detected in bile duct epithelial cells |
| doi_str_mv | 10.1371/journal.pntd.0000398 |
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Here, we describe the cDNA, gene organization, phylogenetic relationships, immunolocalization, and functional characterization of the cathepsin F cysteine protease gene, here termed Ov-cf-1, from O. viverrini. The full length mRNA of 1020 nucleotides (nt) encoded a 326 amino acid zymogen consisting of a predicted signal peptide (18 amino acids, aa), prosegment (95 aa), and mature protease (213 aa). BLAST analysis using the Ov-CF-1 protein as the query revealed that the protease shared identity with cathepsin F-like cysteine proteases of other trematodes, including Clonorchis sinensis (81%), Paragonimus westermani (58%), Schistosoma mansoni and S. japonicum (52%), and with vertebrate cathepsin F (51%). Transcripts encoding the protease were detected in all developmental stages that parasitize the mammalian host. The Ov-cf-1 gene, of approximately 3 kb in length, included seven exons interrupted by six introns; the exons ranged from 69 to 267 bp in length, the introns from 43 to 1,060 bp. The six intron/exon boundaries of Ov-cf-1 were conserved with intron/exon boundaries in the human cathepsin F gene, although the gene structure of human cathepsin F is more complex. Unlike Ov-CF-1, human cathepsin F zymogen includes a cystatin domain in the prosegment region. Phylogenetic analysis revealed that the fluke, human, and other cathepsin Fs branched together in a clade discrete from the cathepsin L cysteine proteases. A recombinant Ov-CF-1 zymogen that displayed low-level activity was expressed in the yeast Pichia pastoris. Although the recombinant protease did not autocatalytically process and activate to a mature enzyme, trans-processing by Fasciola hepatica cathepsin L cleaved the prosegment of Ov-CF-1, releasing a mature cathepsin F with activity against the peptide Z-Phe-Arg-NHMec >50 times that of the zymogen. Immunocytochemistry using antibodies raised against the recombinant enzyme showed that Ov-CF-1 is expressed in the gut of the mature hermaphroditic fluke and also in the reproductive structures, including vitelline glands, egg, and testis. Ov-CF-1 was detected in bile duct epithelial cells surrounding the flukes several weeks after infection of hamsters with O. viverrini and, in addition, had accumulated in the secondary (small) bile ducts where flukes cannot reach due to their large size.
A cathepsin F cysteine protease of the human liver fluke O. viverrini has been characterized at the gene and protein level. Secretion of this protease may contribute to the hepatobiliary abnormalities, including cholangiocarcinogenesis, observed in individuals infected with this parasite.</description><identifier>ISSN: 1935-2735</identifier><identifier>ISSN: 1935-2727</identifier><identifier>EISSN: 1935-2735</identifier><identifier>DOI: 10.1371/journal.pntd.0000398</identifier><identifier>PMID: 19308250</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Amino Acid Sequence ; Animals ; Biochemistry/Bioinformatics ; Cancer ; Carcinogens ; Cathepsin F - genetics ; Cathepsin F - metabolism ; Cricetinae ; Helminth Proteins - genetics ; Helminth Proteins - metabolism ; Humans ; Immunization ; Immunohistochemistry ; Infections ; Liver ; Liver - parasitology ; Molecular Sequence Data ; Opisthorchiasis - parasitology ; Opisthorchis - enzymology ; Opisthorchis - genetics ; Parasites ; Parasitic diseases ; Phylogenetics ; Phylogeny ; Scholarships & fellowships ; Sequence Alignment ; Tropical diseases</subject><ispartof>PLoS neglected tropical diseases, 2009, Vol.3 (3), p.e398</ispartof><rights>2009 Pinlaor et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited: Pinlaor P, Kaewpitoon N, Laha T, Sripa B, Kaewkes S, et al. (2009) Cathepsin F Cysteine Protease of the Human Liver Fluke, Opisthorchis viverrini. PLoS Negl Trop Dis 3(3): e398. doi:10.1371/journal.pntd.0000398</rights><rights>Pinlaor et al. 2009</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c525t-2240b728a2ec9b5e3803f9c881380d43f4a8a9166e7e35bfa4ae65f4b79c9f8d3</citedby><cites>FETCH-LOGICAL-c525t-2240b728a2ec9b5e3803f9c881380d43f4a8a9166e7e35bfa4ae65f4b79c9f8d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2654340/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2654340/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2101,2927,4023,23865,27922,27923,27924,53790,53792,79471,79472</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19308250$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Knox, David</contributor><creatorcontrib>Pinlaor, Porntip</creatorcontrib><creatorcontrib>Kaewpitoon, Natthawut</creatorcontrib><creatorcontrib>Laha, Thewarach</creatorcontrib><creatorcontrib>Sripa, Banchob</creatorcontrib><creatorcontrib>Kaewkes, Sasithorn</creatorcontrib><creatorcontrib>Morales, Maria E</creatorcontrib><creatorcontrib>Mann, Victoria H</creatorcontrib><creatorcontrib>Parriott, Sandi K</creatorcontrib><creatorcontrib>Suttiprapa, Sutas</creatorcontrib><creatorcontrib>Robinson, Mark W</creatorcontrib><creatorcontrib>To, Joyce</creatorcontrib><creatorcontrib>Dalton, John P</creatorcontrib><creatorcontrib>Loukas, Alex</creatorcontrib><creatorcontrib>Brindley, Paul J</creatorcontrib><title>Cathepsin F cysteine protease of the human liver fluke, Opisthorchis viverrini</title><title>PLoS neglected tropical diseases</title><addtitle>PLoS Negl Trop Dis</addtitle><description>The liver fluke Opisthorchis viverrini is classified as a class I carcinogen due to the association between cholangiocarcinoma and chronic O. viverrini infection. During its feeding activity within the bile duct, the parasite secretes several cathepsin F cysteine proteases that may induce or contribute to the pathologies associated with hepatobiliary abnormalities.
Here, we describe the cDNA, gene organization, phylogenetic relationships, immunolocalization, and functional characterization of the cathepsin F cysteine protease gene, here termed Ov-cf-1, from O. viverrini. The full length mRNA of 1020 nucleotides (nt) encoded a 326 amino acid zymogen consisting of a predicted signal peptide (18 amino acids, aa), prosegment (95 aa), and mature protease (213 aa). BLAST analysis using the Ov-CF-1 protein as the query revealed that the protease shared identity with cathepsin F-like cysteine proteases of other trematodes, including Clonorchis sinensis (81%), Paragonimus westermani (58%), Schistosoma mansoni and S. japonicum (52%), and with vertebrate cathepsin F (51%). Transcripts encoding the protease were detected in all developmental stages that parasitize the mammalian host. The Ov-cf-1 gene, of approximately 3 kb in length, included seven exons interrupted by six introns; the exons ranged from 69 to 267 bp in length, the introns from 43 to 1,060 bp. The six intron/exon boundaries of Ov-cf-1 were conserved with intron/exon boundaries in the human cathepsin F gene, although the gene structure of human cathepsin F is more complex. Unlike Ov-CF-1, human cathepsin F zymogen includes a cystatin domain in the prosegment region. Phylogenetic analysis revealed that the fluke, human, and other cathepsin Fs branched together in a clade discrete from the cathepsin L cysteine proteases. A recombinant Ov-CF-1 zymogen that displayed low-level activity was expressed in the yeast Pichia pastoris. Although the recombinant protease did not autocatalytically process and activate to a mature enzyme, trans-processing by Fasciola hepatica cathepsin L cleaved the prosegment of Ov-CF-1, releasing a mature cathepsin F with activity against the peptide Z-Phe-Arg-NHMec >50 times that of the zymogen. Immunocytochemistry using antibodies raised against the recombinant enzyme showed that Ov-CF-1 is expressed in the gut of the mature hermaphroditic fluke and also in the reproductive structures, including vitelline glands, egg, and testis. Ov-CF-1 was detected in bile duct epithelial cells surrounding the flukes several weeks after infection of hamsters with O. viverrini and, in addition, had accumulated in the secondary (small) bile ducts where flukes cannot reach due to their large size.
A cathepsin F cysteine protease of the human liver fluke O. viverrini has been characterized at the gene and protein level. Secretion of this protease may contribute to the hepatobiliary abnormalities, including cholangiocarcinogenesis, observed in individuals infected with this parasite.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Biochemistry/Bioinformatics</subject><subject>Cancer</subject><subject>Carcinogens</subject><subject>Cathepsin F - genetics</subject><subject>Cathepsin F - metabolism</subject><subject>Cricetinae</subject><subject>Helminth Proteins - genetics</subject><subject>Helminth Proteins - metabolism</subject><subject>Humans</subject><subject>Immunization</subject><subject>Immunohistochemistry</subject><subject>Infections</subject><subject>Liver</subject><subject>Liver - parasitology</subject><subject>Molecular Sequence Data</subject><subject>Opisthorchiasis - parasitology</subject><subject>Opisthorchis - enzymology</subject><subject>Opisthorchis - genetics</subject><subject>Parasites</subject><subject>Parasitic diseases</subject><subject>Phylogenetics</subject><subject>Phylogeny</subject><subject>Scholarships & fellowships</subject><subject>Sequence Alignment</subject><subject>Tropical 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cysteine protease of the human liver fluke, Opisthorchis viverrini</title><author>Pinlaor, Porntip ; Kaewpitoon, Natthawut ; Laha, Thewarach ; Sripa, Banchob ; Kaewkes, Sasithorn ; Morales, Maria E ; Mann, Victoria H ; Parriott, Sandi K ; Suttiprapa, Sutas ; Robinson, Mark W ; To, Joyce ; Dalton, John P ; Loukas, Alex ; Brindley, Paul J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c525t-2240b728a2ec9b5e3803f9c881380d43f4a8a9166e7e35bfa4ae65f4b79c9f8d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Biochemistry/Bioinformatics</topic><topic>Cancer</topic><topic>Carcinogens</topic><topic>Cathepsin F - genetics</topic><topic>Cathepsin F - metabolism</topic><topic>Cricetinae</topic><topic>Helminth Proteins - genetics</topic><topic>Helminth Proteins - metabolism</topic><topic>Humans</topic><topic>Immunization</topic><topic>Immunohistochemistry</topic><topic>Infections</topic><topic>Liver</topic><topic>Liver - parasitology</topic><topic>Molecular Sequence Data</topic><topic>Opisthorchiasis - parasitology</topic><topic>Opisthorchis - enzymology</topic><topic>Opisthorchis - genetics</topic><topic>Parasites</topic><topic>Parasitic diseases</topic><topic>Phylogenetics</topic><topic>Phylogeny</topic><topic>Scholarships & fellowships</topic><topic>Sequence Alignment</topic><topic>Tropical diseases</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pinlaor, Porntip</creatorcontrib><creatorcontrib>Kaewpitoon, Natthawut</creatorcontrib><creatorcontrib>Laha, Thewarach</creatorcontrib><creatorcontrib>Sripa, Banchob</creatorcontrib><creatorcontrib>Kaewkes, Sasithorn</creatorcontrib><creatorcontrib>Morales, Maria E</creatorcontrib><creatorcontrib>Mann, Victoria H</creatorcontrib><creatorcontrib>Parriott, 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Dis</addtitle><date>2009</date><risdate>2009</risdate><volume>3</volume><issue>3</issue><spage>e398</spage><pages>e398-</pages><issn>1935-2735</issn><issn>1935-2727</issn><eissn>1935-2735</eissn><abstract>The liver fluke Opisthorchis viverrini is classified as a class I carcinogen due to the association between cholangiocarcinoma and chronic O. viverrini infection. During its feeding activity within the bile duct, the parasite secretes several cathepsin F cysteine proteases that may induce or contribute to the pathologies associated with hepatobiliary abnormalities.
Here, we describe the cDNA, gene organization, phylogenetic relationships, immunolocalization, and functional characterization of the cathepsin F cysteine protease gene, here termed Ov-cf-1, from O. viverrini. The full length mRNA of 1020 nucleotides (nt) encoded a 326 amino acid zymogen consisting of a predicted signal peptide (18 amino acids, aa), prosegment (95 aa), and mature protease (213 aa). BLAST analysis using the Ov-CF-1 protein as the query revealed that the protease shared identity with cathepsin F-like cysteine proteases of other trematodes, including Clonorchis sinensis (81%), Paragonimus westermani (58%), Schistosoma mansoni and S. japonicum (52%), and with vertebrate cathepsin F (51%). Transcripts encoding the protease were detected in all developmental stages that parasitize the mammalian host. The Ov-cf-1 gene, of approximately 3 kb in length, included seven exons interrupted by six introns; the exons ranged from 69 to 267 bp in length, the introns from 43 to 1,060 bp. The six intron/exon boundaries of Ov-cf-1 were conserved with intron/exon boundaries in the human cathepsin F gene, although the gene structure of human cathepsin F is more complex. Unlike Ov-CF-1, human cathepsin F zymogen includes a cystatin domain in the prosegment region. Phylogenetic analysis revealed that the fluke, human, and other cathepsin Fs branched together in a clade discrete from the cathepsin L cysteine proteases. A recombinant Ov-CF-1 zymogen that displayed low-level activity was expressed in the yeast Pichia pastoris. Although the recombinant protease did not autocatalytically process and activate to a mature enzyme, trans-processing by Fasciola hepatica cathepsin L cleaved the prosegment of Ov-CF-1, releasing a mature cathepsin F with activity against the peptide Z-Phe-Arg-NHMec >50 times that of the zymogen. Immunocytochemistry using antibodies raised against the recombinant enzyme showed that Ov-CF-1 is expressed in the gut of the mature hermaphroditic fluke and also in the reproductive structures, including vitelline glands, egg, and testis. Ov-CF-1 was detected in bile duct epithelial cells surrounding the flukes several weeks after infection of hamsters with O. viverrini and, in addition, had accumulated in the secondary (small) bile ducts where flukes cannot reach due to their large size.
A cathepsin F cysteine protease of the human liver fluke O. viverrini has been characterized at the gene and protein level. Secretion of this protease may contribute to the hepatobiliary abnormalities, including cholangiocarcinogenesis, observed in individuals infected with this parasite.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>19308250</pmid><doi>10.1371/journal.pntd.0000398</doi><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1935-2735 |
| ispartof | PLoS neglected tropical diseases, 2009, Vol.3 (3), p.e398 |
| issn | 1935-2735 1935-2727 1935-2735 |
| language | eng |
| recordid | cdi_plos_journals_1288105334 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central Open Access; Public Library of Science (PLoS); PubMed Central |
| subjects | Amino Acid Sequence Animals Biochemistry/Bioinformatics Cancer Carcinogens Cathepsin F - genetics Cathepsin F - metabolism Cricetinae Helminth Proteins - genetics Helminth Proteins - metabolism Humans Immunization Immunohistochemistry Infections Liver Liver - parasitology Molecular Sequence Data Opisthorchiasis - parasitology Opisthorchis - enzymology Opisthorchis - genetics Parasites Parasitic diseases Phylogenetics Phylogeny Scholarships & fellowships Sequence Alignment Tropical diseases |
| title | Cathepsin F cysteine protease of the human liver fluke, Opisthorchis viverrini |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-08T08%3A26%3A25IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Cathepsin%20F%20cysteine%20protease%20of%20the%20human%20liver%20fluke,%20Opisthorchis%20viverrini&rft.jtitle=PLoS%20neglected%20tropical%20diseases&rft.au=Pinlaor,%20Porntip&rft.date=2009&rft.volume=3&rft.issue=3&rft.spage=e398&rft.pages=e398-&rft.issn=1935-2735&rft.eissn=1935-2735&rft_id=info:doi/10.1371/journal.pntd.0000398&rft_dat=%3Cproquest_plos_%3E2893314361%3C/proquest_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1288105334&rft_id=info:pmid/19308250&rft_doaj_id=oai_doaj_org_article_80155b8ade724e52a78524e33dfb3970&rfr_iscdi=true |