Molecular cloning and characterization of cDNA encoding a ubiquitin-conjugating enzyme from Clonorchis sinensis
The ubiquitin-proteasome system is an essential mechanism for protein degradation in eukaryotes. Protein ubiquitination is composed of a series of enzymatic reactions. The ubiquitin-conjugating enzyme (E2) is one of the important enzymes involved in the process. A cDNA encoding an E2 enzyme was clon...
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| Veröffentlicht in: | Parasitology research (1987) 2004-10, Vol.94 (3), p.227-232 |
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| container_title | Parasitology research (1987) |
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| creator | Song, Linxia Chen, Shouyi Yu, Xinbing Wu, Zhongdao Xu, Jin Yang, Guang Zheng, Nancai Hu, Xuchu Guo, Lingchen Dai, Jianfeng Xu, Jian Ji, Chaoneng Gu, Shaohua Ying, Kang |
| description | The ubiquitin-proteasome system is an essential mechanism for protein degradation in eukaryotes. Protein ubiquitination is composed of a series of enzymatic reactions. The ubiquitin-conjugating enzyme (E2) is one of the important enzymes involved in the process. A cDNA encoding an E2 enzyme was cloned from a Clonorchis sinensis cDNA library by large-scale sequencing. This new cDNA contains 862 bp with a putative open reading frame of 156 amino acids. The deduced amino acid sequence is 77% identical to the human E2, HHR6A and HHR6B. The coding region of this cDNA was expressed in E. coli as a GST-tagged protein, and was purified to electrophoretic homogeneity. Enzymatic assays showed that this E2 had the capacity to form a thiolester linkage, and could conjugate ubiquitin to histone H2A in an E3-independent manner in vitro, which indicated that the expressed protein was functionally active. The nucleotide sequence reported in this paper has been submitted to the Genbank Database with accession number AY632078. |
| doi_str_mv | 10.1007/s00436-004-1206-5 |
| format | Article |
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Protein ubiquitination is composed of a series of enzymatic reactions. The ubiquitin-conjugating enzyme (E2) is one of the important enzymes involved in the process. A cDNA encoding an E2 enzyme was cloned from a Clonorchis sinensis cDNA library by large-scale sequencing. This new cDNA contains 862 bp with a putative open reading frame of 156 amino acids. The deduced amino acid sequence is 77% identical to the human E2, HHR6A and HHR6B. The coding region of this cDNA was expressed in E. coli as a GST-tagged protein, and was purified to electrophoretic homogeneity. Enzymatic assays showed that this E2 had the capacity to form a thiolester linkage, and could conjugate ubiquitin to histone H2A in an E3-independent manner in vitro, which indicated that the expressed protein was functionally active. The nucleotide sequence reported in this paper has been submitted to the Genbank Database with accession number AY632078.</description><identifier>ISSN: 0932-0113</identifier><identifier>EISSN: 1432-1955</identifier><identifier>DOI: 10.1007/s00436-004-1206-5</identifier><identifier>PMID: 15480785</identifier><identifier>CODEN: PARREZ</identifier><language>eng</language><publisher>Berlin: Springer</publisher><subject>Amino Acid Sequence ; Animals ; Base Sequence ; Biological and medical sciences ; Cloning, Molecular ; Clonorchis sinensis - enzymology ; Clonorchis sinensis - genetics ; DNA, Complementary - genetics ; DNA, Helminth - genetics ; Fundamental and applied biological sciences. Psychology ; Gene Library ; General aspects ; General aspects and techniques. Study of several systematic groups. Models ; Genes, Helminth ; Humans ; Invertebrates ; Molecular Sequence Data ; Recombinant Proteins - genetics ; Recombinant Proteins - metabolism ; Sequence Homology, Amino Acid ; Ubiquitin-Conjugating Enzymes - genetics ; Ubiquitin-Conjugating Enzymes - metabolism</subject><ispartof>Parasitology research (1987), 2004-10, Vol.94 (3), p.227-232</ispartof><rights>2004 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c259t-33080c6a0b9e6115f5d279f61b10ee994e8566703575e0d8652e4a84d46edf213</citedby><cites>FETCH-LOGICAL-c259t-33080c6a0b9e6115f5d279f61b10ee994e8566703575e0d8652e4a84d46edf213</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=16156522$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15480785$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Song, Linxia</creatorcontrib><creatorcontrib>Chen, Shouyi</creatorcontrib><creatorcontrib>Yu, Xinbing</creatorcontrib><creatorcontrib>Wu, Zhongdao</creatorcontrib><creatorcontrib>Xu, Jin</creatorcontrib><creatorcontrib>Yang, Guang</creatorcontrib><creatorcontrib>Zheng, Nancai</creatorcontrib><creatorcontrib>Hu, Xuchu</creatorcontrib><creatorcontrib>Guo, Lingchen</creatorcontrib><creatorcontrib>Dai, Jianfeng</creatorcontrib><creatorcontrib>Xu, Jian</creatorcontrib><creatorcontrib>Ji, Chaoneng</creatorcontrib><creatorcontrib>Gu, Shaohua</creatorcontrib><creatorcontrib>Ying, Kang</creatorcontrib><title>Molecular cloning and characterization of cDNA encoding a ubiquitin-conjugating enzyme from Clonorchis sinensis</title><title>Parasitology research (1987)</title><addtitle>Parasitol Res</addtitle><description>The ubiquitin-proteasome system is an essential mechanism for protein degradation in eukaryotes. Protein ubiquitination is composed of a series of enzymatic reactions. The ubiquitin-conjugating enzyme (E2) is one of the important enzymes involved in the process. A cDNA encoding an E2 enzyme was cloned from a Clonorchis sinensis cDNA library by large-scale sequencing. This new cDNA contains 862 bp with a putative open reading frame of 156 amino acids. The deduced amino acid sequence is 77% identical to the human E2, HHR6A and HHR6B. The coding region of this cDNA was expressed in E. coli as a GST-tagged protein, and was purified to electrophoretic homogeneity. Enzymatic assays showed that this E2 had the capacity to form a thiolester linkage, and could conjugate ubiquitin to histone H2A in an E3-independent manner in vitro, which indicated that the expressed protein was functionally active. The nucleotide sequence reported in this paper has been submitted to the Genbank Database with accession number AY632078.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Cloning, Molecular</subject><subject>Clonorchis sinensis - enzymology</subject><subject>Clonorchis sinensis - genetics</subject><subject>DNA, Complementary - genetics</subject><subject>DNA, Helminth - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Library</subject><subject>General aspects</subject><subject>General aspects and techniques. Study of several systematic groups. Models</subject><subject>Genes, Helminth</subject><subject>Humans</subject><subject>Invertebrates</subject><subject>Molecular Sequence Data</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - metabolism</subject><subject>Sequence Homology, Amino Acid</subject><subject>Ubiquitin-Conjugating Enzymes - genetics</subject><subject>Ubiquitin-Conjugating Enzymes - metabolism</subject><issn>0932-0113</issn><issn>1432-1955</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkM1O5DAQhK0VaBkGHoAL8gVuYbsT20mOaPiV2N3L7tnyOB0wSmywk8Pw9HiYkbhUt1pflVrF2BnCFQLUvxKAqFSRtcASVCF_sAWKqiywlfKALaDNOyBWR-w4pVcArJUQP9kRStFA3cgFC7_DQHYeTOR2CN75Z258x-2LicZOFN2HmVzwPPTc3vy55uRt6L4oPq_d--wm5wsb_Ov8nMF8J_-xGYn3MYx8lRNDtC8u8eQ8-eTSCTvszZDodD-X7P_d7b_VQ_H09_5xdf1U2FK2U1FV0IBVBtYtKUTZy66s217hGoGobQU1UqkaKllLgq5RsiRhGtEJRV1fYrVkl7vctxjeZ0qTHl2yNAzGU5iTVqpVdalEBnEH2hhSitTrt-hGEzcaQW9b1ruWdVa9bVnL7Dnfh8_rkbpvx77WDFzsAZOsGfpovHXpm1Mo88dl9Qmr34W7</recordid><startdate>20041001</startdate><enddate>20041001</enddate><creator>Song, Linxia</creator><creator>Chen, Shouyi</creator><creator>Yu, Xinbing</creator><creator>Wu, Zhongdao</creator><creator>Xu, Jin</creator><creator>Yang, Guang</creator><creator>Zheng, Nancai</creator><creator>Hu, Xuchu</creator><creator>Guo, Lingchen</creator><creator>Dai, Jianfeng</creator><creator>Xu, Jian</creator><creator>Ji, Chaoneng</creator><creator>Gu, Shaohua</creator><creator>Ying, Kang</creator><general>Springer</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20041001</creationdate><title>Molecular cloning and characterization of cDNA encoding a ubiquitin-conjugating enzyme from Clonorchis sinensis</title><author>Song, Linxia ; Chen, Shouyi ; Yu, Xinbing ; Wu, Zhongdao ; Xu, Jin ; Yang, Guang ; Zheng, Nancai ; Hu, Xuchu ; Guo, Lingchen ; Dai, Jianfeng ; Xu, Jian ; Ji, Chaoneng ; Gu, Shaohua ; Ying, Kang</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c259t-33080c6a0b9e6115f5d279f61b10ee994e8566703575e0d8652e4a84d46edf213</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Cloning, Molecular</topic><topic>Clonorchis sinensis - enzymology</topic><topic>Clonorchis sinensis - genetics</topic><topic>DNA, Complementary - genetics</topic><topic>DNA, Helminth - genetics</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Library</topic><topic>General aspects</topic><topic>General aspects and techniques. Study of several systematic groups. Models</topic><topic>Genes, Helminth</topic><topic>Humans</topic><topic>Invertebrates</topic><topic>Molecular Sequence Data</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - metabolism</topic><topic>Sequence Homology, Amino Acid</topic><topic>Ubiquitin-Conjugating Enzymes - genetics</topic><topic>Ubiquitin-Conjugating Enzymes - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Song, Linxia</creatorcontrib><creatorcontrib>Chen, Shouyi</creatorcontrib><creatorcontrib>Yu, Xinbing</creatorcontrib><creatorcontrib>Wu, Zhongdao</creatorcontrib><creatorcontrib>Xu, Jin</creatorcontrib><creatorcontrib>Yang, Guang</creatorcontrib><creatorcontrib>Zheng, Nancai</creatorcontrib><creatorcontrib>Hu, Xuchu</creatorcontrib><creatorcontrib>Guo, Lingchen</creatorcontrib><creatorcontrib>Dai, Jianfeng</creatorcontrib><creatorcontrib>Xu, Jian</creatorcontrib><creatorcontrib>Ji, Chaoneng</creatorcontrib><creatorcontrib>Gu, Shaohua</creatorcontrib><creatorcontrib>Ying, Kang</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Parasitology research (1987)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Song, Linxia</au><au>Chen, Shouyi</au><au>Yu, Xinbing</au><au>Wu, Zhongdao</au><au>Xu, Jin</au><au>Yang, Guang</au><au>Zheng, Nancai</au><au>Hu, Xuchu</au><au>Guo, Lingchen</au><au>Dai, Jianfeng</au><au>Xu, Jian</au><au>Ji, Chaoneng</au><au>Gu, Shaohua</au><au>Ying, Kang</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular cloning and characterization of cDNA encoding a ubiquitin-conjugating enzyme from Clonorchis sinensis</atitle><jtitle>Parasitology research (1987)</jtitle><addtitle>Parasitol Res</addtitle><date>2004-10-01</date><risdate>2004</risdate><volume>94</volume><issue>3</issue><spage>227</spage><epage>232</epage><pages>227-232</pages><issn>0932-0113</issn><eissn>1432-1955</eissn><coden>PARREZ</coden><abstract>The ubiquitin-proteasome system is an essential mechanism for protein degradation in eukaryotes. Protein ubiquitination is composed of a series of enzymatic reactions. The ubiquitin-conjugating enzyme (E2) is one of the important enzymes involved in the process. A cDNA encoding an E2 enzyme was cloned from a Clonorchis sinensis cDNA library by large-scale sequencing. This new cDNA contains 862 bp with a putative open reading frame of 156 amino acids. The deduced amino acid sequence is 77% identical to the human E2, HHR6A and HHR6B. The coding region of this cDNA was expressed in E. coli as a GST-tagged protein, and was purified to electrophoretic homogeneity. Enzymatic assays showed that this E2 had the capacity to form a thiolester linkage, and could conjugate ubiquitin to histone H2A in an E3-independent manner in vitro, which indicated that the expressed protein was functionally active. The nucleotide sequence reported in this paper has been submitted to the Genbank Database with accession number AY632078.</abstract><cop>Berlin</cop><pub>Springer</pub><pmid>15480785</pmid><doi>10.1007/s00436-004-1206-5</doi><tpages>6</tpages></addata></record> |
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| ispartof | Parasitology research (1987), 2004-10, Vol.94 (3), p.227-232 |
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| subjects | Amino Acid Sequence Animals Base Sequence Biological and medical sciences Cloning, Molecular Clonorchis sinensis - enzymology Clonorchis sinensis - genetics DNA, Complementary - genetics DNA, Helminth - genetics Fundamental and applied biological sciences. Psychology Gene Library General aspects General aspects and techniques. Study of several systematic groups. Models Genes, Helminth Humans Invertebrates Molecular Sequence Data Recombinant Proteins - genetics Recombinant Proteins - metabolism Sequence Homology, Amino Acid Ubiquitin-Conjugating Enzymes - genetics Ubiquitin-Conjugating Enzymes - metabolism |
| title | Molecular cloning and characterization of cDNA encoding a ubiquitin-conjugating enzyme from Clonorchis sinensis |
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