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Title: A small single-domain protein folds through the same pathway on and off the ribosome

Abstract

In vivo, proteins fold and function in a complex environment subject to many stresses that can modulate a protein’s energy landscape. One aspect of the environment pertinent to protein folding is the ribosome, since proteins have the opportunity to fold while still bound to the ribosome during translation. We use a combination of force and chemical denaturant (chemomechanical unfolding), as well as point mutations, to characterize the folding mechanism of the src SH3 domain both as a stalled ribosome nascent chain and free in solution. Our results indicate that src SH3 folds through the same pathway on and off the ribosome. Molecular simulations also indicate that the ribosome does not affect the folding pathway for this small protein. Taken together, we conclude that the ribosome does not alter the folding mechanism of this small protein. These results, if general, suggest the ribosome may exert a bigger influence on the folding of multidomain proteins or protein domains that can partially fold before the entire domain sequence is outside the ribosome exit tunnel

Authors:
 [1];  [2];  [3]; ORCiD logo [2];  [4]
  1. Univ. of California, Berkeley, CA (United States). Inst. for Quantitative Biosciences (QB3)
  2. National Inst. of Health (NIH), Bethesda, MD (United States). National Inst. of Diabetes and Digestive and Kidney Disease, Lab. of Chemical Physics
  3. Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology
  4. Univ. of California, Berkeley, CA (United States). Inst. for Quantitative Biosciences (QB3), Dept. of Molecular and Cell Biology, Dept. of Chemistry
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1561900
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 115; Journal Issue: 48; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; protein folding; ribosome; cotranslational folding; single-molecule force spectroscopy; optical tweezers

Citation Formats

Guinn, Emily J., Tian, Pengfei, Shin, Mia, Best, Robert B., and Marqusee, Susan. A small single-domain protein folds through the same pathway on and off the ribosome. United States: N. p., 2018. Web. doi:10.1073/pnas.1810517115.
Guinn, Emily J., Tian, Pengfei, Shin, Mia, Best, Robert B., & Marqusee, Susan. A small single-domain protein folds through the same pathway on and off the ribosome. United States. doi:10.1073/pnas.1810517115.
Guinn, Emily J., Tian, Pengfei, Shin, Mia, Best, Robert B., and Marqusee, Susan. Thu . "A small single-domain protein folds through the same pathway on and off the ribosome". United States. doi:10.1073/pnas.1810517115. https://www.osti.gov/servlets/purl/1561900.
@article{osti_1561900,
title = {A small single-domain protein folds through the same pathway on and off the ribosome},
author = {Guinn, Emily J. and Tian, Pengfei and Shin, Mia and Best, Robert B. and Marqusee, Susan},
abstractNote = {In vivo, proteins fold and function in a complex environment subject to many stresses that can modulate a protein’s energy landscape. One aspect of the environment pertinent to protein folding is the ribosome, since proteins have the opportunity to fold while still bound to the ribosome during translation. We use a combination of force and chemical denaturant (chemomechanical unfolding), as well as point mutations, to characterize the folding mechanism of the src SH3 domain both as a stalled ribosome nascent chain and free in solution. Our results indicate that src SH3 folds through the same pathway on and off the ribosome. Molecular simulations also indicate that the ribosome does not affect the folding pathway for this small protein. Taken together, we conclude that the ribosome does not alter the folding mechanism of this small protein. These results, if general, suggest the ribosome may exert a bigger influence on the folding of multidomain proteins or protein domains that can partially fold before the entire domain sequence is outside the ribosome exit tunnel},
doi = {10.1073/pnas.1810517115},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 48,
volume = 115,
place = {United States},
year = {2018},
month = {11}
}

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