Amino acid substitution at the substrate-binding subsite alters the specificity of the Phanerochaete chrysosporium cellobiose dehydrogenase.
Identifieur interne : 000590 ( Main/Exploration ); précédent : 000589; suivant : 000591Amino acid substitution at the substrate-binding subsite alters the specificity of the Phanerochaete chrysosporium cellobiose dehydrogenase.
Auteurs : Desriani [Japon] ; Stefano Ferri ; Koji SodeSource :
- Biochemical and biophysical research communications [ 1090-2104 ] ; 2010.
Descripteurs français
- KwdFr :
- Carbohydrate dehydrogenases (composition chimique), Carbohydrate dehydrogenases (génétique), Carbohydrate dehydrogenases (métabolisme), Flavines (métabolisme), Lactose (métabolisme), Phanerochaete (enzymologie), Sites de fixation (génétique), Spécificité du substrat (génétique), Structure tertiaire des protéines (MeSH), Substitution d'acide aminé (MeSH).
- MESH :
- composition chimique : Carbohydrate dehydrogenases.
- enzymologie : Phanerochaete.
- génétique : Carbohydrate dehydrogenases, Sites de fixation, Spécificité du substrat.
- métabolisme : Carbohydrate dehydrogenases, Flavines, Lactose.
- Structure tertiaire des protéines, Substitution d'acide aminé.
English descriptors
- KwdEn :
- Amino Acid Substitution (MeSH), Binding Sites (genetics), Carbohydrate Dehydrogenases (chemistry), Carbohydrate Dehydrogenases (genetics), Carbohydrate Dehydrogenases (metabolism), Flavins (metabolism), Lactose (metabolism), Phanerochaete (enzymology), Protein Structure, Tertiary (MeSH), Substrate Specificity (genetics).
- MESH :
- chemical , chemistry : Carbohydrate Dehydrogenases.
- enzymology : Phanerochaete.
- genetics : Binding Sites, Carbohydrate Dehydrogenases, Substrate Specificity.
- chemical , metabolism : Carbohydrate Dehydrogenases, Flavins, Lactose.
- Amino Acid Substitution, Protein Structure, Tertiary.
Abstract
The active site of cellobiose dehydrogenase from Phanerochaete chrysosporium is composed of two subsites, a catalytic C subsite and a substrate-binding B subsite. Based on the crystal structure of the enzyme with a cellobiose analogue, residue Glu279 was selected for site-directed mutagenesis studies. Substitution of Glu279 to Ala, Asn, and Asp had no effect on the expression of the protein in Pichia pastoris but completely abolished its enzymatic activity. Substitution of Glu279 to Gln drastically altered the enzyme's substrate specificity. While the wild-type cellobiose dehydrogenase efficiently oxidizes cellobiose and lactose, the Glu279Gln mutant retained most of its activity with cellobiose but was completely inactive with lactose. We generated structural models of the active site interacting with cellobiose and lactose to provide an interpretation of these results.
DOI: 10.1016/j.bbrc.2009.12.052
PubMed: 20120044
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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(enzymologie)</term><term>Sites de fixation (génétique)</term><term>Spécificité du substrat (génétique)</term><term>Structure tertiaire des protéines (MeSH)</term><term>Substitution d'acide aminé (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Carbohydrate Dehydrogenases</term></keywords><keywords scheme="MESH" qualifier="composition chimique" xml:lang="fr"><term>Carbohydrate dehydrogenases</term></keywords><keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Phanerochaete</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Phanerochaete</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Binding Sites</term><term>Carbohydrate Dehydrogenases</term><term>Substrate Specificity</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Carbohydrate dehydrogenases</term><term>Sites de fixation</term><term>Spécificité du 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Based on the crystal structure of the enzyme with a cellobiose analogue, residue Glu279 was selected for site-directed mutagenesis studies. Substitution of Glu279 to Ala, Asn, and Asp had no effect on the expression of the protein in Pichia pastoris but completely abolished its enzymatic activity. Substitution of Glu279 to Gln drastically altered the enzyme's substrate specificity. While the wild-type cellobiose dehydrogenase efficiently oxidizes cellobiose and lactose, the Glu279Gln mutant retained most of its activity with cellobiose but was completely inactive with lactose. We generated structural models of the active site interacting with cellobiose and lactose to provide an interpretation of these results.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">20120044</PMID><DateCompleted><Year>2010</Year><Month>03</Month><Day>08</Day></DateCompleted><DateRevised><Year>2019</Year><Month>06</Month><Day>12</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Electronic">1090-2104</ISSN><JournalIssue CitedMedium="Internet"><Volume>391</Volume><Issue>2</Issue><PubDate><Year>2010</Year><Month>Jan</Month><Day>08</Day></PubDate></JournalIssue><Title>Biochemical and biophysical research communications</Title><ISOAbbreviation>Biochem Biophys Res Commun</ISOAbbreviation></Journal><ArticleTitle>Amino acid substitution at the substrate-binding subsite alters the specificity of the Phanerochaete chrysosporium cellobiose dehydrogenase.</ArticleTitle><Pagination><MedlinePgn>1246-50</MedlinePgn></Pagination><Abstract><AbstractText>The active site of cellobiose dehydrogenase from Phanerochaete chrysosporium is composed of two subsites, a catalytic C subsite and a substrate-binding B subsite. Based on the crystal structure of the enzyme with a cellobiose analogue, residue Glu279 was selected for site-directed mutagenesis studies. Substitution of Glu279 to Ala, Asn, and Asp had no effect on the expression of the protein in Pichia pastoris but completely abolished its enzymatic activity. Substitution of Glu279 to Gln drastically altered the enzyme's substrate specificity. While the wild-type cellobiose dehydrogenase efficiently oxidizes cellobiose and lactose, the Glu279Gln mutant retained most of its activity with cellobiose but was completely inactive with lactose. 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