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Tracing a protein’s folding pathway over evolutionary time using ancestral sequence reconstruction and hydrogen exchange

The conformations populated during protein folding have been studied for decades; yet, their evolutionary importance remains largely unexplored. Ancestral sequence reconstruction allows access to proteins across evolutionary time, and new methods such as pulsed-labeling hydrogen exchange coupled wit...

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Detalles Bibliográficos
Autores principales: Lim, Shion An, Bolin, Eric Richard, Marqusee, Susan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6158009/
https://www.ncbi.nlm.nih.gov/pubmed/30204082
http://dx.doi.org/10.7554/eLife.38369
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author Lim, Shion An
Bolin, Eric Richard
Marqusee, Susan
author_facet Lim, Shion An
Bolin, Eric Richard
Marqusee, Susan
author_sort Lim, Shion An
collection PubMed
description The conformations populated during protein folding have been studied for decades; yet, their evolutionary importance remains largely unexplored. Ancestral sequence reconstruction allows access to proteins across evolutionary time, and new methods such as pulsed-labeling hydrogen exchange coupled with mass spectrometry allow determination of folding intermediate structures at near amino-acid resolution. Here, we combine these techniques to monitor the folding of the ribonuclease H family along the evolutionary lineages of T. thermophilus and E. coli RNase H. All homologs and ancestral proteins studied populate a similar folding intermediate despite being separated by billions of years of evolution. Even though this conformation is conserved, the pathway leading to it has diverged over evolutionary time, and rational mutations can alter this trajectory. Our results demonstrate that evolutionary processes can affect the energy landscape to preserve or alter specific features of a protein’s folding pathway.
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spelling pubmed-61580092018-09-27 Tracing a protein’s folding pathway over evolutionary time using ancestral sequence reconstruction and hydrogen exchange Lim, Shion An Bolin, Eric Richard Marqusee, Susan eLife Biochemistry and Chemical Biology The conformations populated during protein folding have been studied for decades; yet, their evolutionary importance remains largely unexplored. Ancestral sequence reconstruction allows access to proteins across evolutionary time, and new methods such as pulsed-labeling hydrogen exchange coupled with mass spectrometry allow determination of folding intermediate structures at near amino-acid resolution. Here, we combine these techniques to monitor the folding of the ribonuclease H family along the evolutionary lineages of T. thermophilus and E. coli RNase H. All homologs and ancestral proteins studied populate a similar folding intermediate despite being separated by billions of years of evolution. Even though this conformation is conserved, the pathway leading to it has diverged over evolutionary time, and rational mutations can alter this trajectory. Our results demonstrate that evolutionary processes can affect the energy landscape to preserve or alter specific features of a protein’s folding pathway. eLife Sciences Publications, Ltd 2018-09-11 /pmc/articles/PMC6158009/ /pubmed/30204082 http://dx.doi.org/10.7554/eLife.38369 Text en © 2018, Lim et al http://creativecommons.org/licenses/by/4.0/ 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 Biochemistry and Chemical Biology
Lim, Shion An
Bolin, Eric Richard
Marqusee, Susan
Tracing a protein’s folding pathway over evolutionary time using ancestral sequence reconstruction and hydrogen exchange
title Tracing a protein’s folding pathway over evolutionary time using ancestral sequence reconstruction and hydrogen exchange
title_full Tracing a protein’s folding pathway over evolutionary time using ancestral sequence reconstruction and hydrogen exchange
title_fullStr Tracing a protein’s folding pathway over evolutionary time using ancestral sequence reconstruction and hydrogen exchange
title_full_unstemmed Tracing a protein’s folding pathway over evolutionary time using ancestral sequence reconstruction and hydrogen exchange
title_short Tracing a protein’s folding pathway over evolutionary time using ancestral sequence reconstruction and hydrogen exchange
title_sort tracing a protein’s folding pathway over evolutionary time using ancestral sequence reconstruction and hydrogen exchange
topic Biochemistry and Chemical Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6158009/
https://www.ncbi.nlm.nih.gov/pubmed/30204082
http://dx.doi.org/10.7554/eLife.38369
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