Cloning, high level-expression and characterization of human lens thioltransferase.
Identifieur interne : 001147 ( Main/Exploration ); précédent : 001146; suivant : 001148Cloning, high level-expression and characterization of human lens thioltransferase.
Auteurs : N. Raghavachari [États-Unis] ; F. Qiao ; T. Shinohara ; T. Kikuchi ; M F LouSource :
- Experimental eye research [ 0014-4835 ] ; 1998.
Descripteurs français
- KwdFr :
- Cellules cultivées (MeSH), Clones cellulaires (MeSH), Cristallin (enzymologie), Cristallines (métabolisme), Données de séquences moléculaires (MeSH), Glutarédoxines (MeSH), Humains (MeSH), Oxidoreductases (métabolisme), Protein-disulfide reductase (glutathione) (MeSH), Réaction de polymérisation en chaîne (MeSH), Séquence d'acides aminés (MeSH), Séquence nucléotidique (MeSH), Technique de Northern (MeSH), Technique de Southern (MeSH).
- MESH :
- enzymologie : Cristallin.
- métabolisme : Cristallines, Oxidoreductases.
- Cellules cultivées, Clones cellulaires, Données de séquences moléculaires, Glutarédoxines, Humains, Protein-disulfide reductase (glutathione), Réaction de polymérisation en chaîne, Séquence d'acides aminés, Séquence nucléotidique, Technique de Northern, Technique de Southern.
English descriptors
- KwdEn :
- Amino Acid Sequence (MeSH), Base Sequence (MeSH), Blotting, Northern (MeSH), Blotting, Southern (MeSH), Cells, Cultured (MeSH), Clone Cells (MeSH), Crystallins (metabolism), Glutaredoxins (MeSH), Humans (MeSH), Lens, Crystalline (enzymology), Molecular Sequence Data (MeSH), Oxidoreductases (metabolism), Polymerase Chain Reaction (MeSH), Protein Disulfide Reductase (Glutathione) (MeSH).
- MESH :
- chemical , metabolism : Crystallins, Oxidoreductases.
- enzymology : Lens, Crystalline.
- Amino Acid Sequence, Base Sequence, Blotting, Northern, Blotting, Southern, Cells, Cultured, Clone Cells, Glutaredoxins, Humans, Molecular Sequence Data, Polymerase Chain Reaction, Protein Disulfide Reductase (Glutathione).
Abstract
Polymerase chain reaction (PCR) primers, directed against the nucleotide sequence of pig liver thioltransferase (PLTT) were used to amplify human lens thioltransferase (HLTT) from a pool of human lens cDNA. The 520 bp PCR fragment obtained was cloned unidirectionally into pCR 3.1-Uni vector and sequenced. The cDNA sequence of the lens thioltransferase had 98% and 87% homology to pig liver and human placental thioltransferases (TTase) respectively. Nhe1 and EcoR1 fragment of the recombinant PCR 3.1-Uni vector was subcloned in pET 23a Expression vector. High level expression of HLTT was accomplished in Escherichia coli and the expressed protein was characterized by immunoblot analysis with anti PLTT and N-terminal amino acid sequence analysis. The recombinant enzyme efficiently dethiolated protein thiol mixed disulfides conjugated to both cystine (PSSC) and glutathione (PSSG) and had a significant dehydroascorbate reductase activity. Human lens thioltransferase thus displayed structural and functional characteristics identical to pig liver and human placental thioltransferases.
DOI: 10.1006/exer.1997.0449
PubMed: 9593639
Affiliations:
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Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Cloning, high level-expression and characterization of human lens thioltransferase.</title><author><name sortKey="Raghavachari, N" sort="Raghavachari, N" uniqKey="Raghavachari N" first="N" last="Raghavachari">N. Raghavachari</name><affiliation wicri:level="1"><nlm:affiliation>Center for Biotechnology, Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, NE, 68583-0905, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Center for Biotechnology, Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, NE, 68583-0905</wicri:regionArea><wicri:noRegion>68583-0905</wicri:noRegion></affiliation></author><author><name sortKey="Qiao, F" sort="Qiao, F" uniqKey="Qiao F" first="F" last="Qiao">F. Qiao</name></author><author><name sortKey="Shinohara, T" sort="Shinohara, T" uniqKey="Shinohara T" first="T" last="Shinohara">T. Shinohara</name></author><author><name sortKey="Kikuchi, T" sort="Kikuchi, T" uniqKey="Kikuchi T" first="T" last="Kikuchi">T. Kikuchi</name></author><author><name sortKey="Lou, M F" sort="Lou, M F" uniqKey="Lou M" first="M F" last="Lou">M F Lou</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="1998">1998</date><idno type="RBID">pubmed:9593639</idno><idno type="pmid">9593639</idno><idno type="doi">10.1006/exer.1997.0449</idno><idno type="wicri:Area/Main/Corpus">001142</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">001142</idno><idno type="wicri:Area/Main/Curation">001142</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">001142</idno><idno type="wicri:Area/Main/Exploration">001142</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Cloning, high level-expression and characterization of human lens thioltransferase.</title><author><name sortKey="Raghavachari, N" sort="Raghavachari, N" uniqKey="Raghavachari N" first="N" last="Raghavachari">N. Raghavachari</name><affiliation wicri:level="1"><nlm:affiliation>Center for Biotechnology, Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, NE, 68583-0905, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Center for Biotechnology, Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, NE, 68583-0905</wicri:regionArea><wicri:noRegion>68583-0905</wicri:noRegion></affiliation></author><author><name sortKey="Qiao, F" sort="Qiao, F" uniqKey="Qiao F" first="F" last="Qiao">F. Qiao</name></author><author><name sortKey="Shinohara, T" sort="Shinohara, T" uniqKey="Shinohara T" first="T" last="Shinohara">T. Shinohara</name></author><author><name sortKey="Kikuchi, T" sort="Kikuchi, T" uniqKey="Kikuchi T" first="T" last="Kikuchi">T. Kikuchi</name></author><author><name sortKey="Lou, M F" sort="Lou, M F" uniqKey="Lou M" first="M F" last="Lou">M F Lou</name></author></analytic><series><title level="j">Experimental eye research</title><idno type="ISSN">0014-4835</idno><imprint><date when="1998" type="published">1998</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence (MeSH)</term><term>Base Sequence (MeSH)</term><term>Blotting, Northern (MeSH)</term><term>Blotting, Southern (MeSH)</term><term>Cells, Cultured (MeSH)</term><term>Clone Cells (MeSH)</term><term>Crystallins (metabolism)</term><term>Glutaredoxins (MeSH)</term><term>Humans (MeSH)</term><term>Lens, Crystalline (enzymology)</term><term>Molecular Sequence Data (MeSH)</term><term>Oxidoreductases (metabolism)</term><term>Polymerase Chain Reaction (MeSH)</term><term>Protein Disulfide Reductase (Glutathione) (MeSH)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Cellules cultivées (MeSH)</term><term>Clones cellulaires (MeSH)</term><term>Cristallin (enzymologie)</term><term>Cristallines (métabolisme)</term><term>Données de séquences moléculaires (MeSH)</term><term>Glutarédoxines (MeSH)</term><term>Humains (MeSH)</term><term>Oxidoreductases (métabolisme)</term><term>Protein-disulfide reductase (glutathione) (MeSH)</term><term>Réaction de polymérisation en chaîne (MeSH)</term><term>Séquence d'acides aminés (MeSH)</term><term>Séquence nucléotidique (MeSH)</term><term>Technique de Northern (MeSH)</term><term>Technique de Southern (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Crystallins</term><term>Oxidoreductases</term></keywords><keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Cristallin</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Lens, Crystalline</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Cristallines</term><term>Oxidoreductases</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Base Sequence</term><term>Blotting, Northern</term><term>Blotting, Southern</term><term>Cells, Cultured</term><term>Clone Cells</term><term>Glutaredoxins</term><term>Humans</term><term>Molecular Sequence Data</term><term>Polymerase Chain Reaction</term><term>Protein Disulfide Reductase (Glutathione)</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Cellules cultivées</term><term>Clones cellulaires</term><term>Données de séquences moléculaires</term><term>Glutarédoxines</term><term>Humains</term><term>Protein-disulfide reductase (glutathione)</term><term>Réaction de polymérisation en chaîne</term><term>Séquence d'acides aminés</term><term>Séquence nucléotidique</term><term>Technique de Northern</term><term>Technique de Southern</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Polymerase chain reaction (PCR) primers, directed against the nucleotide sequence of pig liver thioltransferase (PLTT) were used to amplify human lens thioltransferase (HLTT) from a pool of human lens cDNA. The 520 bp PCR fragment obtained was cloned unidirectionally into pCR 3.1-Uni vector and sequenced. The cDNA sequence of the lens thioltransferase had 98% and 87% homology to pig liver and human placental thioltransferases (TTase) respectively. Nhe1 and EcoR1 fragment of the recombinant PCR 3.1-Uni vector was subcloned in pET 23a Expression vector. High level expression of HLTT was accomplished in Escherichia coli and the expressed protein was characterized by immunoblot analysis with anti PLTT and N-terminal amino acid sequence analysis. The recombinant enzyme efficiently dethiolated protein thiol mixed disulfides conjugated to both cystine (PSSC) and glutathione (PSSG) and had a significant dehydroascorbate reductase activity. Human lens thioltransferase thus displayed structural and functional characteristics identical to pig liver and human placental thioltransferases.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">9593639</PMID><DateCompleted><Year>1998</Year><Month>07</Month><Day>31</Day></DateCompleted><DateRevised><Year>2007</Year><Month>11</Month><Day>15</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0014-4835</ISSN><JournalIssue CitedMedium="Print"><Volume>66</Volume><Issue>4</Issue><PubDate><Year>1998</Year><Month>Apr</Month></PubDate></JournalIssue><Title>Experimental eye research</Title><ISOAbbreviation>Exp Eye Res</ISOAbbreviation></Journal><ArticleTitle>Cloning, high level-expression and characterization of human lens thioltransferase.</ArticleTitle><Pagination><MedlinePgn>465-75</MedlinePgn></Pagination><Abstract><AbstractText>Polymerase chain reaction (PCR) primers, directed against the nucleotide sequence of pig liver thioltransferase (PLTT) were used to amplify human lens thioltransferase (HLTT) from a pool of human lens cDNA. The 520 bp PCR fragment obtained was cloned unidirectionally into pCR 3.1-Uni vector and sequenced. The cDNA sequence of the lens thioltransferase had 98% and 87% homology to pig liver and human placental thioltransferases (TTase) respectively. Nhe1 and EcoR1 fragment of the recombinant PCR 3.1-Uni vector was subcloned in pET 23a Expression vector. High level expression of HLTT was accomplished in Escherichia coli and the expressed protein was characterized by immunoblot analysis with anti PLTT and N-terminal amino acid sequence analysis. The recombinant enzyme efficiently dethiolated protein thiol mixed disulfides conjugated to both cystine (PSSC) and glutathione (PSSG) and had a significant dehydroascorbate reductase activity. Human lens thioltransferase thus displayed structural and functional characteristics identical to pig liver and human placental thioltransferases.</AbstractText><CopyrightInformation>Copyright 1998 Academic Press Limited.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Raghavachari</LastName><ForeName>N</ForeName><Initials>N</Initials><AffiliationInfo><Affiliation>Center for Biotechnology, Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, NE, 68583-0905, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Qiao</LastName><ForeName>F</ForeName><Initials>F</Initials></Author><Author ValidYN="Y"><LastName>Shinohara</LastName><ForeName>T</ForeName><Initials>T</Initials></Author><Author ValidYN="Y"><LastName>Kikuchi</LastName><ForeName>T</ForeName><Initials>T</Initials></Author><Author ValidYN="Y"><LastName>Lou</LastName><ForeName>M F</ForeName><Initials>MF</Initials></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>EY 06647-01</GrantID><Acronym>EY</Acronym><Agency>NEI NIH HHS</Agency><Country>United States</Country></Grant><Grant><GrantID>EY 10595</GrantID><Acronym>EY</Acronym><Agency>NEI NIH HHS</Agency><Country>United States</Country></Grant></GrantList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013487">Research Support, U.S. Gov't, P.H.S.</PublicationType><PublicationType UI="D016441">Retracted Publication</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Exp Eye Res</MedlineTA><NlmUniqueID>0370707</NlmUniqueID><ISSNLinking>0014-4835</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D003459">Crystallins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D054477">Glutaredoxins</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 1.-</RegistryNumber><NameOfSubstance UI="D010088">Oxidoreductases</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 1.8.4.2</RegistryNumber><NameOfSubstance UI="D011490">Protein Disulfide Reductase (Glutathione)</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 1.8.5.1</RegistryNumber><NameOfSubstance UI="C020666">glutathione dehydrogenase (ascorbate)</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><CommentsCorrectionsList><CommentsCorrections RefType="RetractionIn"><RefSource>Raghavachari N, Qiao F, Shinohara T, Kikuchi T, Lou MF. Exp Eye Res. 1999 Nov;69(5):583</RefSource><PMID Version="1">10712076</PMID></CommentsCorrections></CommentsCorrectionsList><MeshHeadingList><MeshHeading><DescriptorName UI="D000595" MajorTopicYN="N">Amino Acid Sequence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D001483" MajorTopicYN="N">Base Sequence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D015152" MajorTopicYN="N">Blotting, Northern</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D015139" MajorTopicYN="N">Blotting, Southern</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002478" MajorTopicYN="N">Cells, Cultured</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002999" MajorTopicYN="N">Clone Cells</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D003459" MajorTopicYN="N">Crystallins</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D054477" MajorTopicYN="N">Glutaredoxins</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D007908" MajorTopicYN="N">Lens, Crystalline</DescriptorName><QualifierName UI="Q000201" MajorTopicYN="Y">enzymology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D008969" MajorTopicYN="N">Molecular Sequence Data</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D010088" MajorTopicYN="N">Oxidoreductases</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D016133" MajorTopicYN="N">Polymerase Chain Reaction</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D011490" MajorTopicYN="Y">Protein Disulfide Reductase (Glutathione)</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>1998</Year><Month>6</Month><Day>17</Day><Hour>2</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2000</Year><Month>3</Month><Day>11</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>1998</Year><Month>6</Month><Day>17</Day><Hour>2</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">9593639</ArticleId><ArticleId IdType="pii">S0014-4835(97)90449-8</ArticleId><ArticleId IdType="doi">10.1006/exer.1997.0449</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>États-Unis</li></country></list><tree><noCountry><name sortKey="Kikuchi, T" sort="Kikuchi, T" uniqKey="Kikuchi T" first="T" last="Kikuchi">T. Kikuchi</name><name sortKey="Lou, M F" sort="Lou, M F" uniqKey="Lou M" first="M F" last="Lou">M F Lou</name><name sortKey="Qiao, F" sort="Qiao, F" uniqKey="Qiao F" first="F" last="Qiao">F. Qiao</name><name sortKey="Shinohara, T" sort="Shinohara, T" uniqKey="Shinohara T" first="T" last="Shinohara">T. Shinohara</name></noCountry><country name="États-Unis"><noRegion><name sortKey="Raghavachari, N" sort="Raghavachari, N" uniqKey="Raghavachari N" first="N" last="Raghavachari">N. Raghavachari</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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