N-terminus deletion affecting the preparation of soluble cyanobacterial glutaredoxin in Escherichia coli.
Identifieur interne : 000C73 ( Main/Exploration ); précédent : 000C72; suivant : 000C74N-terminus deletion affecting the preparation of soluble cyanobacterial glutaredoxin in Escherichia coli.
Auteurs : Min Li [République populaire de Chine] ; Qing Yang ; Yang Gao ; Qingyu WuSource :
- Biochemistry. Biokhimiia [ 0006-2979 ] ; 2007.
Descripteurs français
- KwdFr :
- Catalyse (MeSH), Cyanobactéries (génétique), Cyanobactéries (métabolisme), Dichroïsme circulaire (MeSH), Données de séquences moléculaires (MeSH), Délétion de séquence (MeSH), Escherichia coli (génétique), Glutarédoxines (MeSH), Interactions hydrophobes et hydrophiles (MeSH), Mutation (MeSH), Oxidoreductases (composition chimique), Oxidoreductases (génétique), Oxidoreductases (métabolisme), Protéines bactériennes (composition chimique), Protéines bactériennes (génétique), Protéines bactériennes (métabolisme), Protéines recombinantes (composition chimique), Protéines recombinantes (isolement et purification), Protéines recombinantes (métabolisme), Similitude de séquences d'acides aminés (MeSH), Solubilité (MeSH), Spectrométrie de fluorescence (MeSH), Stress oxydatif (MeSH), Séquence d'acides aminés (MeSH), Électrophorèse sur gel de polyacrylamide (MeSH).
- MESH :
- composition chimique : Oxidoreductases, Protéines bactériennes, Protéines recombinantes.
- génétique : Cyanobactéries, Escherichia coli, Oxidoreductases, Protéines bactériennes.
- isolement et purification : Protéines recombinantes.
- métabolisme : Cyanobactéries, Oxidoreductases, Protéines bactériennes, Protéines recombinantes.
- Catalyse, Dichroïsme circulaire, Données de séquences moléculaires, Délétion de séquence, Glutarédoxines, Interactions hydrophobes et hydrophiles, Mutation, Similitude de séquences d'acides aminés, Solubilité, Spectrométrie de fluorescence, Stress oxydatif, Séquence d'acides aminés, Électrophorèse sur gel de polyacrylamide.
English descriptors
- KwdEn :
- Amino Acid Sequence (MeSH), Bacterial Proteins (chemistry), Bacterial Proteins (genetics), Bacterial Proteins (metabolism), Catalysis (MeSH), Circular Dichroism (MeSH), Cyanobacteria (genetics), Cyanobacteria (metabolism), Electrophoresis, Polyacrylamide Gel (MeSH), Escherichia coli (genetics), Glutaredoxins (MeSH), Hydrophobic and Hydrophilic Interactions (MeSH), Molecular Sequence Data (MeSH), Mutation (MeSH), Oxidative Stress (MeSH), Oxidoreductases (chemistry), Oxidoreductases (genetics), Oxidoreductases (metabolism), Recombinant Proteins (chemistry), Recombinant Proteins (isolation & purification), Recombinant Proteins (metabolism), Sequence Deletion (MeSH), Sequence Homology, Amino Acid (MeSH), Solubility (MeSH), Spectrometry, Fluorescence (MeSH).
- MESH :
- chemical , chemistry : Bacterial Proteins, Oxidoreductases, Recombinant Proteins.
- chemical , genetics : Bacterial Proteins, Oxidoreductases.
- chemical , isolation & purification : Recombinant Proteins.
- chemical , metabolism : Bacterial Proteins, Oxidoreductases, Recombinant Proteins.
- genetics : Cyanobacteria, Escherichia coli.
- metabolism : Cyanobacteria.
- Amino Acid Sequence, Catalysis, Circular Dichroism, Electrophoresis, Polyacrylamide Gel, Glutaredoxins, Hydrophobic and Hydrophilic Interactions, Molecular Sequence Data, Mutation, Oxidative Stress, Sequence Deletion, Sequence Homology, Amino Acid, Solubility, Spectrometry, Fluorescence.
Abstract
Glutaredoxin has been implicated in maintenance of a normal cellular thiol/disufide ratio and the regeneration of oxidatively damaged proteins. In order to obtain more information about these important regulatory proteins in cyanobacteria, we have previously cloned and expressed the first cyanobacterial glutaredoxin gene ssr2061 in Escherichia coli. In this work, the second glutaredoxin gene slr1562 was studied. About 90% of Grx2061 coded by ssr2061 was produced in a soluble form while 90% of Grx1562 coded by slr1562 was found in inclusion bodies. To improve the production of soluble Grx1562, we constructed two mutants: Grx1562NC with cysteines in conserved site substituted by serines, and Grx1562M with N-terminus hydrophobic region deletion. Only the latter mutant was successfully expressed in soluble form with increased glutaredoxin activity and showed less sensitivity in oxidative stress. Spectroscopic analysis shows that the structure of Grx1562M with less hydrophobic nature could give more opportunity for protein solubility and could improve the substrate catalytic efficiency. These results suggest that hydrophobic N-terminus determines the insolubility of Grx1562 and may provide another strategy for increasing expression level of soluble heterologous proteins in E. coli.
DOI: 10.1134/s0006297907030091
PubMed: 17447885
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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Biokhimiia</title><idno type="ISSN">0006-2979</idno><imprint><date when="2007" type="published">2007</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence (MeSH)</term><term>Bacterial Proteins (chemistry)</term><term>Bacterial Proteins (genetics)</term><term>Bacterial Proteins (metabolism)</term><term>Catalysis (MeSH)</term><term>Circular Dichroism (MeSH)</term><term>Cyanobacteria (genetics)</term><term>Cyanobacteria (metabolism)</term><term>Electrophoresis, Polyacrylamide Gel (MeSH)</term><term>Escherichia coli (genetics)</term><term>Glutaredoxins (MeSH)</term><term>Hydrophobic and Hydrophilic Interactions (MeSH)</term><term>Molecular Sequence Data (MeSH)</term><term>Mutation (MeSH)</term><term>Oxidative Stress (MeSH)</term><term>Oxidoreductases (chemistry)</term><term>Oxidoreductases (genetics)</term><term>Oxidoreductases (metabolism)</term><term>Recombinant Proteins (chemistry)</term><term>Recombinant Proteins (isolation & purification)</term><term>Recombinant Proteins (metabolism)</term><term>Sequence Deletion (MeSH)</term><term>Sequence Homology, Amino Acid (MeSH)</term><term>Solubility (MeSH)</term><term>Spectrometry, Fluorescence (MeSH)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Catalyse (MeSH)</term><term>Cyanobactéries (génétique)</term><term>Cyanobactéries (métabolisme)</term><term>Dichroïsme circulaire (MeSH)</term><term>Données de séquences moléculaires (MeSH)</term><term>Délétion de séquence (MeSH)</term><term>Escherichia coli (génétique)</term><term>Glutarédoxines (MeSH)</term><term>Interactions hydrophobes et hydrophiles (MeSH)</term><term>Mutation (MeSH)</term><term>Oxidoreductases (composition chimique)</term><term>Oxidoreductases (génétique)</term><term>Oxidoreductases (métabolisme)</term><term>Protéines bactériennes (composition chimique)</term><term>Protéines bactériennes (génétique)</term><term>Protéines bactériennes (métabolisme)</term><term>Protéines recombinantes (composition chimique)</term><term>Protéines recombinantes (isolement et purification)</term><term>Protéines recombinantes (métabolisme)</term><term>Similitude de séquences d'acides aminés (MeSH)</term><term>Solubilité (MeSH)</term><term>Spectrométrie de fluorescence (MeSH)</term><term>Stress oxydatif (MeSH)</term><term>Séquence d'acides aminés (MeSH)</term><term>Électrophorèse sur gel de polyacrylamide (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Bacterial Proteins</term><term>Oxidoreductases</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Bacterial Proteins</term><term>Oxidoreductases</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Bacterial Proteins</term><term>Oxidoreductases</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" qualifier="composition chimique" xml:lang="fr"><term>Oxidoreductases</term><term>Protéines bactériennes</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Cyanobacteria</term><term>Escherichia coli</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Cyanobactéries</term><term>Escherichia coli</term><term>Oxidoreductases</term><term>Protéines bactériennes</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Cyanobacteria</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Cyanobactéries</term><term>Oxidoreductases</term><term>Protéines bactériennes</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Catalysis</term><term>Circular Dichroism</term><term>Electrophoresis, Polyacrylamide Gel</term><term>Glutaredoxins</term><term>Hydrophobic and Hydrophilic Interactions</term><term>Molecular Sequence Data</term><term>Mutation</term><term>Oxidative Stress</term><term>Sequence Deletion</term><term>Sequence Homology, Amino Acid</term><term>Solubility</term><term>Spectrometry, Fluorescence</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Catalyse</term><term>Dichroïsme circulaire</term><term>Données de séquences moléculaires</term><term>Délétion de séquence</term><term>Glutarédoxines</term><term>Interactions hydrophobes et hydrophiles</term><term>Mutation</term><term>Similitude de séquences d'acides aminés</term><term>Solubilité</term><term>Spectrométrie de fluorescence</term><term>Stress oxydatif</term><term>Séquence d'acides aminés</term><term>Électrophorèse sur gel de polyacrylamide</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Glutaredoxin has been implicated in maintenance of a normal cellular thiol/disufide ratio and the regeneration of oxidatively damaged proteins. In order to obtain more information about these important regulatory proteins in cyanobacteria, we have previously cloned and expressed the first cyanobacterial glutaredoxin gene ssr2061 in Escherichia coli. In this work, the second glutaredoxin gene slr1562 was studied. About 90% of Grx2061 coded by ssr2061 was produced in a soluble form while 90% of Grx1562 coded by slr1562 was found in inclusion bodies. To improve the production of soluble Grx1562, we constructed two mutants: Grx1562NC with cysteines in conserved site substituted by serines, and Grx1562M with N-terminus hydrophobic region deletion. Only the latter mutant was successfully expressed in soluble form with increased glutaredoxin activity and showed less sensitivity in oxidative stress. Spectroscopic analysis shows that the structure of Grx1562M with less hydrophobic nature could give more opportunity for protein solubility and could improve the substrate catalytic efficiency. These results suggest that hydrophobic N-terminus determines the insolubility of Grx1562 and may provide another strategy for increasing expression level of soluble heterologous proteins in E. coli.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">17447885</PMID><DateCompleted><Year>2007</Year><Month>08</Month><Day>06</Day></DateCompleted><DateRevised><Year>2019</Year><Month>09</Month><Day>17</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0006-2979</ISSN><JournalIssue CitedMedium="Print"><Volume>72</Volume><Issue>3</Issue><PubDate><Year>2007</Year><Month>Mar</Month></PubDate></JournalIssue><Title>Biochemistry. Biokhimiia</Title><ISOAbbreviation>Biochemistry (Mosc)</ISOAbbreviation></Journal><ArticleTitle>N-terminus deletion affecting the preparation of soluble cyanobacterial glutaredoxin in Escherichia coli.</ArticleTitle><Pagination><MedlinePgn>313-9</MedlinePgn></Pagination><Abstract><AbstractText>Glutaredoxin has been implicated in maintenance of a normal cellular thiol/disufide ratio and the regeneration of oxidatively damaged proteins. In order to obtain more information about these important regulatory proteins in cyanobacteria, we have previously cloned and expressed the first cyanobacterial glutaredoxin gene ssr2061 in Escherichia coli. In this work, the second glutaredoxin gene slr1562 was studied. About 90% of Grx2061 coded by ssr2061 was produced in a soluble form while 90% of Grx1562 coded by slr1562 was found in inclusion bodies. To improve the production of soluble Grx1562, we constructed two mutants: Grx1562NC with cysteines in conserved site substituted by serines, and Grx1562M with N-terminus hydrophobic region deletion. Only the latter mutant was successfully expressed in soluble form with increased glutaredoxin activity and showed less sensitivity in oxidative stress. Spectroscopic analysis shows that the structure of Grx1562M with less hydrophobic nature could give more opportunity for protein solubility and could improve the substrate catalytic efficiency. These results suggest that hydrophobic N-terminus determines the insolubility of Grx1562 and may provide another strategy for increasing expression level of soluble heterologous proteins in E. coli.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Li</LastName><ForeName>Min</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing, PR China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Yang</LastName><ForeName>Qing</ForeName><Initials>Q</Initials></Author><Author ValidYN="Y"><LastName>Gao</LastName><ForeName>Yang</ForeName><Initials>Y</Initials></Author><Author ValidYN="Y"><LastName>Wu</LastName><ForeName>Qingyu</ForeName><Initials>Q</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Biochemistry (Mosc)</MedlineTA><NlmUniqueID>0376536</NlmUniqueID><ISSNLinking>0006-2979</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D001426">Bacterial Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D054477">Glutaredoxins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 1.-</RegistryNumber><NameOfSubstance UI="D010088">Oxidoreductases</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000595" MajorTopicYN="N">Amino Acid Sequence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D001426" MajorTopicYN="N">Bacterial Proteins</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D002384" MajorTopicYN="N">Catalysis</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002942" MajorTopicYN="N">Circular Dichroism</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D000458" MajorTopicYN="N">Cyanobacteria</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004591" MajorTopicYN="N">Electrophoresis, Polyacrylamide Gel</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004926" MajorTopicYN="N">Escherichia coli</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D054477" MajorTopicYN="N">Glutaredoxins</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D057927" MajorTopicYN="N">Hydrophobic and Hydrophilic Interactions</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008969" MajorTopicYN="N">Molecular Sequence Data</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D009154" MajorTopicYN="N">Mutation</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018384" MajorTopicYN="N">Oxidative Stress</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D010088" MajorTopicYN="N">Oxidoreductases</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011994" MajorTopicYN="N">Recombinant Proteins</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000302" MajorTopicYN="N">isolation & purification</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D017384" MajorTopicYN="Y">Sequence Deletion</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D017386" MajorTopicYN="N">Sequence Homology, Amino Acid</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D012995" MajorTopicYN="N">Solubility</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D013050" MajorTopicYN="N">Spectrometry, Fluorescence</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2007</Year><Month>4</Month><Day>24</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2007</Year><Month>8</Month><Day>7</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2007</Year><Month>4</Month><Day>24</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">17447885</ArticleId><ArticleId IdType="pii">BCM72030383</ArticleId><ArticleId IdType="doi">10.1134/s0006297907030091</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>République populaire de Chine</li></country><settlement><li>Pékin</li></settlement></list><tree><noCountry><name sortKey="Gao, Yang" sort="Gao, Yang" uniqKey="Gao Y" first="Yang" last="Gao">Yang Gao</name><name sortKey="Wu, Qingyu" sort="Wu, Qingyu" uniqKey="Wu Q" first="Qingyu" last="Wu">Qingyu Wu</name><name sortKey="Yang, Qing" sort="Yang, Qing" uniqKey="Yang Q" first="Qing" last="Yang">Qing Yang</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Li, Min" sort="Li, Min" uniqKey="Li M" first="Min" last="Li">Min Li</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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