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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...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2018
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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. |
format | Online Article Text |
id | pubmed-6158009 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
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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