Computational site-directed mutagenesis studies of the role of the hydrophobic triad on substrate binding in cholesterol oxidase
© 2017 Wiley Periodicals, Inc. Cholesterol oxidase (ChOx) is a flavoenzyme that oxidizes and isomerizes cholesterol (CHL) to form cholest-4-en-3-one. Molecular docking and molecular dynamics simulations were conducted to predict the binding interactions of CHL in the active site. Several key intera...
| Main Authors: | , , , |
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| Format: | Journal Article |
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John Wiley & Sons, Inc.
2017
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| Online Access: | http://hdl.handle.net/20.500.11937/63291 |
| _version_ | 1848761047039606784 |
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| author | Harb, L. Arooj, Mahreen Vrielink, A. Mancera, Ricardo |
| author_facet | Harb, L. Arooj, Mahreen Vrielink, A. Mancera, Ricardo |
| author_sort | Harb, L. |
| building | Curtin Institutional Repository |
| collection | Online Access |
| description | © 2017 Wiley Periodicals, Inc. Cholesterol oxidase (ChOx) is a flavoenzyme that oxidizes and isomerizes cholesterol (CHL) to form cholest-4-en-3-one. Molecular docking and molecular dynamics simulations were conducted to predict the binding interactions of CHL in the active site. Several key interactions (E361-CHL, N485-FAD, and H447-CHL) were identified and which are likely to determine the correct positioning of CHL relative to flavin-adenine dinucleotide (FAD). Binding of CHL also induced changes in key residues of the active site leading to the closure of the oxygen channel. A group of residues, Y107, F444, and Y446, known as the hydrophobic triad, are believed to affect the binding of CHL in the active site. Computational site-directed mutagenesis of these residues revealed that their mutation affects the conformations of key residues in the active site, leading to non-optimal binding of CHL and to changes in the structure of the oxygen channel, all of which are likely to reduce the catalytic efficiency of ChOx. Proteins 2017; 85:1645–1655. © 2017 Wiley Periodicals, Inc. |
| first_indexed | 2025-11-14T10:25:27Z |
| format | Journal Article |
| id | curtin-20.500.11937-63291 |
| institution | Curtin University Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-14T10:25:27Z |
| publishDate | 2017 |
| publisher | John Wiley & Sons, Inc. |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | curtin-20.500.11937-632912018-02-06T06:24:17Z Computational site-directed mutagenesis studies of the role of the hydrophobic triad on substrate binding in cholesterol oxidase Harb, L. Arooj, Mahreen Vrielink, A. Mancera, Ricardo © 2017 Wiley Periodicals, Inc. Cholesterol oxidase (ChOx) is a flavoenzyme that oxidizes and isomerizes cholesterol (CHL) to form cholest-4-en-3-one. Molecular docking and molecular dynamics simulations were conducted to predict the binding interactions of CHL in the active site. Several key interactions (E361-CHL, N485-FAD, and H447-CHL) were identified and which are likely to determine the correct positioning of CHL relative to flavin-adenine dinucleotide (FAD). Binding of CHL also induced changes in key residues of the active site leading to the closure of the oxygen channel. A group of residues, Y107, F444, and Y446, known as the hydrophobic triad, are believed to affect the binding of CHL in the active site. Computational site-directed mutagenesis of these residues revealed that their mutation affects the conformations of key residues in the active site, leading to non-optimal binding of CHL and to changes in the structure of the oxygen channel, all of which are likely to reduce the catalytic efficiency of ChOx. Proteins 2017; 85:1645–1655. © 2017 Wiley Periodicals, Inc. 2017 Journal Article http://hdl.handle.net/20.500.11937/63291 10.1002/prot.25319 John Wiley & Sons, Inc. restricted |
| spellingShingle | Harb, L. Arooj, Mahreen Vrielink, A. Mancera, Ricardo Computational site-directed mutagenesis studies of the role of the hydrophobic triad on substrate binding in cholesterol oxidase |
| title | Computational site-directed mutagenesis studies of the role of the hydrophobic triad on substrate binding in cholesterol oxidase |
| title_full | Computational site-directed mutagenesis studies of the role of the hydrophobic triad on substrate binding in cholesterol oxidase |
| title_fullStr | Computational site-directed mutagenesis studies of the role of the hydrophobic triad on substrate binding in cholesterol oxidase |
| title_full_unstemmed | Computational site-directed mutagenesis studies of the role of the hydrophobic triad on substrate binding in cholesterol oxidase |
| title_short | Computational site-directed mutagenesis studies of the role of the hydrophobic triad on substrate binding in cholesterol oxidase |
| title_sort | computational site-directed mutagenesis studies of the role of the hydrophobic triad on substrate binding in cholesterol oxidase |
| url | http://hdl.handle.net/20.500.11937/63291 |