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Chemical Mapping Exposes the Importance of Active Site Interactions in Governing the Temperature Dependence of Enzyme Turnover

[Image: see text] Uncovering the role of global protein dynamics in enzyme turnover is needed to fully understand enzyme catalysis. Recently, we have demonstrated that the heat capacity of catalysis, ΔC(P)(‡), can reveal links between the protein free energy landscape, global protein dynamics, and e...

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Autores principales: Winter, Samuel D., Jones, Hannah B. L., Răsădean, Dora M., Crean, Rory M., Danson, Michael J., Pantoş, G. Dan, Katona, Gergely, Prentice, Erica, Arcus, Vickery L., van der Kamp, Marc W., Pudney, Christopher R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2021
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8689651/
https://www.ncbi.nlm.nih.gov/pubmed/34956689
http://dx.doi.org/10.1021/acscatal.1c04679
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author Winter, Samuel D.
Jones, Hannah B. L.
Răsădean, Dora M.
Crean, Rory M.
Danson, Michael J.
Pantoş, G. Dan
Katona, Gergely
Prentice, Erica
Arcus, Vickery L.
van der Kamp, Marc W.
Pudney, Christopher R.
author_facet Winter, Samuel D.
Jones, Hannah B. L.
Răsădean, Dora M.
Crean, Rory M.
Danson, Michael J.
Pantoş, G. Dan
Katona, Gergely
Prentice, Erica
Arcus, Vickery L.
van der Kamp, Marc W.
Pudney, Christopher R.
author_sort Winter, Samuel D.
collection PubMed
description [Image: see text] Uncovering the role of global protein dynamics in enzyme turnover is needed to fully understand enzyme catalysis. Recently, we have demonstrated that the heat capacity of catalysis, ΔC(P)(‡), can reveal links between the protein free energy landscape, global protein dynamics, and enzyme turnover, suggesting that subtle changes in molecular interactions at the active site can affect long-range protein dynamics and link to enzyme temperature activity. Here, we use a model promiscuous enzyme (glucose dehydrogenase from Sulfolobus solfataricus) to chemically map how individual substrate interactions affect the temperature dependence of enzyme activity and the network of motions throughout the protein. Utilizing a combination of kinetics, red edge excitation shift (REES) spectroscopy, and computational simulation, we explore the complex relationship between enzyme–substrate interactions and the global dynamics of the protein. We find that changes in ΔC(P)(‡) and protein dynamics can be mapped to specific substrate–enzyme interactions. Our study reveals how subtle changes in substrate binding affect global changes in motion and flexibility extending throughout the protein.
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spelling pubmed-86896512021-12-22 Chemical Mapping Exposes the Importance of Active Site Interactions in Governing the Temperature Dependence of Enzyme Turnover Winter, Samuel D. Jones, Hannah B. L. Răsădean, Dora M. Crean, Rory M. Danson, Michael J. Pantoş, G. Dan Katona, Gergely Prentice, Erica Arcus, Vickery L. van der Kamp, Marc W. Pudney, Christopher R. ACS Catal [Image: see text] Uncovering the role of global protein dynamics in enzyme turnover is needed to fully understand enzyme catalysis. Recently, we have demonstrated that the heat capacity of catalysis, ΔC(P)(‡), can reveal links between the protein free energy landscape, global protein dynamics, and enzyme turnover, suggesting that subtle changes in molecular interactions at the active site can affect long-range protein dynamics and link to enzyme temperature activity. Here, we use a model promiscuous enzyme (glucose dehydrogenase from Sulfolobus solfataricus) to chemically map how individual substrate interactions affect the temperature dependence of enzyme activity and the network of motions throughout the protein. Utilizing a combination of kinetics, red edge excitation shift (REES) spectroscopy, and computational simulation, we explore the complex relationship between enzyme–substrate interactions and the global dynamics of the protein. We find that changes in ΔC(P)(‡) and protein dynamics can be mapped to specific substrate–enzyme interactions. Our study reveals how subtle changes in substrate binding affect global changes in motion and flexibility extending throughout the protein. American Chemical Society 2021-11-29 2021-12-17 /pmc/articles/PMC8689651/ /pubmed/34956689 http://dx.doi.org/10.1021/acscatal.1c04679 Text en © 2021 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Winter, Samuel D.
Jones, Hannah B. L.
Răsădean, Dora M.
Crean, Rory M.
Danson, Michael J.
Pantoş, G. Dan
Katona, Gergely
Prentice, Erica
Arcus, Vickery L.
van der Kamp, Marc W.
Pudney, Christopher R.
Chemical Mapping Exposes the Importance of Active Site Interactions in Governing the Temperature Dependence of Enzyme Turnover
title Chemical Mapping Exposes the Importance of Active Site Interactions in Governing the Temperature Dependence of Enzyme Turnover
title_full Chemical Mapping Exposes the Importance of Active Site Interactions in Governing the Temperature Dependence of Enzyme Turnover
title_fullStr Chemical Mapping Exposes the Importance of Active Site Interactions in Governing the Temperature Dependence of Enzyme Turnover
title_full_unstemmed Chemical Mapping Exposes the Importance of Active Site Interactions in Governing the Temperature Dependence of Enzyme Turnover
title_short Chemical Mapping Exposes the Importance of Active Site Interactions in Governing the Temperature Dependence of Enzyme Turnover
title_sort chemical mapping exposes the importance of active site interactions in governing the temperature dependence of enzyme turnover
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8689651/
https://www.ncbi.nlm.nih.gov/pubmed/34956689
http://dx.doi.org/10.1021/acscatal.1c04679
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