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Structure of a low-population intermediate state in the release of an enzyme product
Enzymes can increase the rate of biomolecular reactions by several orders of magnitude. Although the steps of substrate capture and product release are essential in the enzymatic process, complete atomic-level descriptions of these steps are difficult to obtain because of the transient nature of the...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4383205/ https://www.ncbi.nlm.nih.gov/pubmed/25575179 http://dx.doi.org/10.7554/eLife.02777 |
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author | De Simone, Alfonso Aprile, Francesco A Dhulesia, Anne Dobson, Christopher M Vendruscolo, Michele |
author_facet | De Simone, Alfonso Aprile, Francesco A Dhulesia, Anne Dobson, Christopher M Vendruscolo, Michele |
author_sort | De Simone, Alfonso |
collection | PubMed |
description | Enzymes can increase the rate of biomolecular reactions by several orders of magnitude. Although the steps of substrate capture and product release are essential in the enzymatic process, complete atomic-level descriptions of these steps are difficult to obtain because of the transient nature of the intermediate conformations, which makes them largely inaccessible to standard structure determination methods. We describe here the determination of the structure of a low-population intermediate in the product release process by human lysozyme through a combination of NMR spectroscopy and molecular dynamics simulations. We validate this structure by rationally designing two mutations, the first engineered to destabilise the intermediate and the second to stabilise it, thus slowing down or speeding up, respectively, product release. These results illustrate how product release by an enzyme can be facilitated by the presence of a metastable intermediate with transient weak interactions between the enzyme and product. DOI: http://dx.doi.org/10.7554/eLife.02777.001 |
format | Online Article Text |
id | pubmed-4383205 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-43832052015-04-03 Structure of a low-population intermediate state in the release of an enzyme product De Simone, Alfonso Aprile, Francesco A Dhulesia, Anne Dobson, Christopher M Vendruscolo, Michele eLife Biophysics and Structural Biology Enzymes can increase the rate of biomolecular reactions by several orders of magnitude. Although the steps of substrate capture and product release are essential in the enzymatic process, complete atomic-level descriptions of these steps are difficult to obtain because of the transient nature of the intermediate conformations, which makes them largely inaccessible to standard structure determination methods. We describe here the determination of the structure of a low-population intermediate in the product release process by human lysozyme through a combination of NMR spectroscopy and molecular dynamics simulations. We validate this structure by rationally designing two mutations, the first engineered to destabilise the intermediate and the second to stabilise it, thus slowing down or speeding up, respectively, product release. These results illustrate how product release by an enzyme can be facilitated by the presence of a metastable intermediate with transient weak interactions between the enzyme and product. DOI: http://dx.doi.org/10.7554/eLife.02777.001 eLife Sciences Publications, Ltd 2015-01-09 /pmc/articles/PMC4383205/ /pubmed/25575179 http://dx.doi.org/10.7554/eLife.02777 Text en © 2014, De Simone et al http://creativecommons.org/licenses/by/4.0/ This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Biophysics and Structural Biology De Simone, Alfonso Aprile, Francesco A Dhulesia, Anne Dobson, Christopher M Vendruscolo, Michele Structure of a low-population intermediate state in the release of an enzyme product |
title | Structure of a low-population intermediate state in the release of an
enzyme product |
title_full | Structure of a low-population intermediate state in the release of an
enzyme product |
title_fullStr | Structure of a low-population intermediate state in the release of an
enzyme product |
title_full_unstemmed | Structure of a low-population intermediate state in the release of an
enzyme product |
title_short | Structure of a low-population intermediate state in the release of an
enzyme product |
title_sort | structure of a low-population intermediate state in the release of an
enzyme product |
topic | Biophysics and Structural Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4383205/ https://www.ncbi.nlm.nih.gov/pubmed/25575179 http://dx.doi.org/10.7554/eLife.02777 |
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