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Title: Unfolding bovine α-lactalbumin with T-jump: Characterizing disordered intermediates via time-resolved x-ray solution scattering and molecular dynamics simulations

Abstract

The protein folding process often proceeds through partially folded transient states. Therefore, a structural understanding of these disordered states is crucial for developing mechanistic models of the folding process. Characterization of unfolded states remains challenging due to their disordered nature, and incorporating multiple methods is necessary. Combining the time-resolved x-ray solution scattering (TRXSS) signal with molecular dynamics (MD), we are able to characterize transient partially folded states of bovine α-lactalbumin, a model system widely used for investigation of molten globule states, during its unfolding triggered by a temperature jump. We track the unfolding process between 20 μs and 70 ms and demonstrate that it passes through three distinct kinetic states. The scattering signals associated with these transient species are then analyzed with TRXSS constrained MD simulations to produce protein structures that are compatible with the input signals. Without utilizing any experimentally extracted kinetic information, the constrained MD simulation successfully drove the protein to an intermediate molten globule state; signals for two later disordered states are refined to terminal unfolded states. From our examination of the structural characteristics of these disordered states, we discuss the implications disordered states have on the folding process, especially on the folding pathway. Finally, we discussmore » the potential applications and limitations of this method.« less

Authors:
ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [2]; ORCiD logo [1]; ORCiD logo [3]
+ Show Author Affiliations
  1. Northwestern Univ., Evanston, IL (United States). Dept. of Chemistry
  2. Univ. of Chicago, IL (United States). Center for Advanced Radiation Sources (CARS)
  3. Northwestern Univ., Evanston, IL (United States). Dept. of Chemistry; Argonne National Lab. (ANL), Argonne, IL (United States). Chemical Sciences and Engineering Division
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE; National Institutes of Health (NIH)
OSTI Identifier:
1813292
Grant/Contract Number:  
AC02-06CH11357; R01-GM115761; 5T32GM008382; SC0000989
Resource Type:
Accepted Manuscript
Journal Name:
Journal of Chemical Physics
Additional Journal Information:
Journal Volume: 154; Journal Issue: 10; Journal ID: ISSN 0021-9606
Publisher:
American Institute of Physics (AIP)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; protein structure; molecular dynamics; protein folding; accessible surface area; transient states; x-ray scattering; folding pathways; metadynamics; statistical thermodynamics

Citation Formats

Hsu, Darren J., Leshchev, Denis, Kosheleva, Irina, Kohlstedt, Kevin L., and Chen, Lin X. Unfolding bovine α-lactalbumin with T-jump: Characterizing disordered intermediates via time-resolved x-ray solution scattering and molecular dynamics simulations. United States: N. p., 2021. Web. doi:10.1063/5.0039194.
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Hsu, Darren J., Leshchev, Denis, Kosheleva, Irina, Kohlstedt, Kevin L., & Chen, Lin X. Unfolding bovine α-lactalbumin with T-jump: Characterizing disordered intermediates via time-resolved x-ray solution scattering and molecular dynamics simulations. United States. https://doi.org/10.1063/5.0039194
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Hsu, Darren J., Leshchev, Denis, Kosheleva, Irina, Kohlstedt, Kevin L., and Chen, Lin X. Sun . "Unfolding bovine α-lactalbumin with T-jump: Characterizing disordered intermediates via time-resolved x-ray solution scattering and molecular dynamics simulations". United States. https://doi.org/10.1063/5.0039194. https://www.osti.gov/servlets/purl/1813292.
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@article{osti_1813292,
title = {Unfolding bovine α-lactalbumin with T-jump: Characterizing disordered intermediates via time-resolved x-ray solution scattering and molecular dynamics simulations},
author = {Hsu, Darren J. and Leshchev, Denis and Kosheleva, Irina and Kohlstedt, Kevin L. and Chen, Lin X.},
abstractNote = {The protein folding process often proceeds through partially folded transient states. Therefore, a structural understanding of these disordered states is crucial for developing mechanistic models of the folding process. Characterization of unfolded states remains challenging due to their disordered nature, and incorporating multiple methods is necessary. Combining the time-resolved x-ray solution scattering (TRXSS) signal with molecular dynamics (MD), we are able to characterize transient partially folded states of bovine α-lactalbumin, a model system widely used for investigation of molten globule states, during its unfolding triggered by a temperature jump. We track the unfolding process between 20 μs and 70 ms and demonstrate that it passes through three distinct kinetic states. The scattering signals associated with these transient species are then analyzed with TRXSS constrained MD simulations to produce protein structures that are compatible with the input signals. Without utilizing any experimentally extracted kinetic information, the constrained MD simulation successfully drove the protein to an intermediate molten globule state; signals for two later disordered states are refined to terminal unfolded states. From our examination of the structural characteristics of these disordered states, we discuss the implications disordered states have on the folding process, especially on the folding pathway. Finally, we discuss the potential applications and limitations of this method.},
doi = {10.1063/5.0039194},
journal = {Journal of Chemical Physics},
number = 10,
volume = 154,
place = {United States},
year = {Sun Mar 14 00:00:00 EST 2021},
month = {Sun Mar 14 00:00:00 EST 2021}
}
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