Temperature-Jump Fluorescence Provides Evidence for Fully Reversible Microsecond Dynamics in a Thermophilic Alcohol Dehydrogenase

[Image: see text] Protein dynamics on the microsecond (μs) time scale were investigated by temperature-jump fluorescence spectroscopy as a function of temperature in two variants of a thermophilic alcohol dehydrogenase: W87F and W87F:H43A. Both mutants exhibit a fast, temperature-independent μs decr...

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Autores principales: Meadows, Corey W., Balakrishnan, Gurusamy, Kier, Brandon L., Spiro, Thomas G., Klinman, Judith P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2015
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4970856/
https://www.ncbi.nlm.nih.gov/pubmed/26223665
http://dx.doi.org/10.1021/jacs.5b04413
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author Meadows, Corey W.
Balakrishnan, Gurusamy
Kier, Brandon L.
Spiro, Thomas G.
Klinman, Judith P.
author_facet Meadows, Corey W.
Balakrishnan, Gurusamy
Kier, Brandon L.
Spiro, Thomas G.
Klinman, Judith P.
author_sort Meadows, Corey W.
collection PubMed
description [Image: see text] Protein dynamics on the microsecond (μs) time scale were investigated by temperature-jump fluorescence spectroscopy as a function of temperature in two variants of a thermophilic alcohol dehydrogenase: W87F and W87F:H43A. Both mutants exhibit a fast, temperature-independent μs decrease in fluorescence followed by a slower full recovery of the initial fluorescence. The results, which rule out an ionizing histidine as the origin of the fluorescence quenching, are discussed in the context of a Trp49-containing dimer interface that acts as a conduit for thermally activated structural change within the protein interior.
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spelling pubmed-49708562016-08-03 Temperature-Jump Fluorescence Provides Evidence for Fully Reversible Microsecond Dynamics in a Thermophilic Alcohol Dehydrogenase Meadows, Corey W. Balakrishnan, Gurusamy Kier, Brandon L. Spiro, Thomas G. Klinman, Judith P. J Am Chem Soc [Image: see text] Protein dynamics on the microsecond (μs) time scale were investigated by temperature-jump fluorescence spectroscopy as a function of temperature in two variants of a thermophilic alcohol dehydrogenase: W87F and W87F:H43A. Both mutants exhibit a fast, temperature-independent μs decrease in fluorescence followed by a slower full recovery of the initial fluorescence. The results, which rule out an ionizing histidine as the origin of the fluorescence quenching, are discussed in the context of a Trp49-containing dimer interface that acts as a conduit for thermally activated structural change within the protein interior. American Chemical Society 2015-07-29 2015-08-19 /pmc/articles/PMC4970856/ /pubmed/26223665 http://dx.doi.org/10.1021/jacs.5b04413 Text en Copyright © 2015 American Chemical Society This is an open access article published under an ACS AuthorChoice License (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) , which permits copying and redistribution of the article or any adaptations for non-commercial purposes.
spellingShingle Meadows, Corey W.
Balakrishnan, Gurusamy
Kier, Brandon L.
Spiro, Thomas G.
Klinman, Judith P.
Temperature-Jump Fluorescence Provides Evidence for Fully Reversible Microsecond Dynamics in a Thermophilic Alcohol Dehydrogenase
title Temperature-Jump Fluorescence Provides Evidence for Fully Reversible Microsecond Dynamics in a Thermophilic Alcohol Dehydrogenase
title_full Temperature-Jump Fluorescence Provides Evidence for Fully Reversible Microsecond Dynamics in a Thermophilic Alcohol Dehydrogenase
title_fullStr Temperature-Jump Fluorescence Provides Evidence for Fully Reversible Microsecond Dynamics in a Thermophilic Alcohol Dehydrogenase
title_full_unstemmed Temperature-Jump Fluorescence Provides Evidence for Fully Reversible Microsecond Dynamics in a Thermophilic Alcohol Dehydrogenase
title_short Temperature-Jump Fluorescence Provides Evidence for Fully Reversible Microsecond Dynamics in a Thermophilic Alcohol Dehydrogenase
title_sort temperature-jump fluorescence provides evidence for fully reversible microsecond dynamics in a thermophilic alcohol dehydrogenase
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4970856/
https://www.ncbi.nlm.nih.gov/pubmed/26223665
http://dx.doi.org/10.1021/jacs.5b04413
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