Probing Cytochrome c Folding Transitions upon Phototriggered Environmental Perturbations Using Time-Resolved X-ray Scattering
Abstract
Direct tracking of protein structural dynamics during folding–unfolding processes is important for understanding the roles of hierarchic structural factors in the formation of functional proteins. As such, using cytochrome c (cyt c) as a platform, we investigated its structural dynamics during folding processes triggered by local environmental changes (i.e., pH or heme iron center oxidation/spin/ligation states) with time-resolved X-ray solution scattering measurements. Starting from partially unfolded cyt c, a sudden pH drop initiated by light excitation of a photoacid caused a structural contraction in microseconds, followed by active site restructuring and unfolding in milliseconds. In contrast, the reduction of iron in the heme via photoinduced electron transfer did not affect conformational stability at short timescales (<1 ms), despite active site coordination geometry changes. These results demonstrate how different environmental perturbations can change the nature of interaction between the active site and protein conformation, even within the same metalloprotein, which will subsequently affect the folding structural dynamics.
- Authors:
-
- Northwestern Univ., Evanston, IL (United States)
- Univ. of Delaware, Newark, DE (United States)
- Univ. of Chicago, IL (United States). Center for Advanced Radiation Sources (CARS)
- Northwestern Univ., Evanston, IL (United States); Argonne National Lab. (ANL), Argonne, IL (United States)
- Publication Date:
- Research Org.:
- Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES); National Institutes of Health (NIH); National Institute of General Medical Sciences (NIGMS); National Institute of Diabetes, Digestive and Kidney Diseases (NIDDK); Northwestern University; E.I. DuPont de Nemours & Co.; The Dow Chemical Company; National Science Foundation (NSF)
- OSTI Identifier:
- 1439630
- Grant/Contract Number:
- R01-GM115761; SC0014664; SC0016288; AC02-06CH11357; R24GM111072; 0960140
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Journal of Physical Chemistry. B, Condensed Matter, Materials, Surfaces, Interfaces and Biophysical Chemistry
- Additional Journal Information:
- Journal Volume: 122; Journal Issue: 20; Journal ID: ISSN 1520-6106
- Publisher:
- American Chemical Society
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; Iron; Structural dynamics; Bioinorganic chemistry; Scattering; Ligation
Citation Formats
Rimmerman, Dolev, Leshchev, Denis, Hsu, Darren J., Hong, Jiyun, Abraham, Baxter, Henning, Robert, Kosheleva, Irina, and Chen, Lin X. Probing Cytochrome c Folding Transitions upon Phototriggered Environmental Perturbations Using Time-Resolved X-ray Scattering. United States: N. p., 2018.
Web. doi:10.1021/acs.jpcb.8b03354.
Rimmerman, Dolev, Leshchev, Denis, Hsu, Darren J., Hong, Jiyun, Abraham, Baxter, Henning, Robert, Kosheleva, Irina, & Chen, Lin X. Probing Cytochrome c Folding Transitions upon Phototriggered Environmental Perturbations Using Time-Resolved X-ray Scattering. United States. https://doi.org/10.1021/acs.jpcb.8b03354
Rimmerman, Dolev, Leshchev, Denis, Hsu, Darren J., Hong, Jiyun, Abraham, Baxter, Henning, Robert, Kosheleva, Irina, and Chen, Lin X. Mon .
"Probing Cytochrome c Folding Transitions upon Phototriggered Environmental Perturbations Using Time-Resolved X-ray Scattering". United States. https://doi.org/10.1021/acs.jpcb.8b03354. https://www.osti.gov/servlets/purl/1439630.
@article{osti_1439630,
title = {Probing Cytochrome c Folding Transitions upon Phototriggered Environmental Perturbations Using Time-Resolved X-ray Scattering},
author = {Rimmerman, Dolev and Leshchev, Denis and Hsu, Darren J. and Hong, Jiyun and Abraham, Baxter and Henning, Robert and Kosheleva, Irina and Chen, Lin X.},
abstractNote = {Direct tracking of protein structural dynamics during folding–unfolding processes is important for understanding the roles of hierarchic structural factors in the formation of functional proteins. As such, using cytochrome c (cyt c) as a platform, we investigated its structural dynamics during folding processes triggered by local environmental changes (i.e., pH or heme iron center oxidation/spin/ligation states) with time-resolved X-ray solution scattering measurements. Starting from partially unfolded cyt c, a sudden pH drop initiated by light excitation of a photoacid caused a structural contraction in microseconds, followed by active site restructuring and unfolding in milliseconds. In contrast, the reduction of iron in the heme via photoinduced electron transfer did not affect conformational stability at short timescales (<1 ms), despite active site coordination geometry changes. These results demonstrate how different environmental perturbations can change the nature of interaction between the active site and protein conformation, even within the same metalloprotein, which will subsequently affect the folding structural dynamics.},
doi = {10.1021/acs.jpcb.8b03354},
journal = {Journal of Physical Chemistry. B, Condensed Matter, Materials, Surfaces, Interfaces and Biophysical Chemistry},
number = 20,
volume = 122,
place = {United States},
year = {2018},
month = {4}
}
Web of Science
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- Photochemical & Photobiological Sciences, Vol. 17, Issue 7
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journal, January 2019
- Hsu, Darren J.; Leshchev, Denis; Rimmerman, Dolev
- Chemical Science, Vol. 10, Issue 42
Photoreduction of ferricytochrome c in the presence of potassium ferrocyanide
journal, January 2018
- Malyshka, Dmitry; Schweitzer-Stenner, Reinhard
- Photochemical & Photobiological Sciences, Vol. 17, Issue 10
Resolving structures of transition metal complex reaction intermediates with femtosecond EXAFS
journal, January 2020
- Britz, Alexander; Abraham, Baxter; Biasin, Elisa
- Physical Chemistry Chemical Physics, Vol. 22, Issue 5