Combinatorial design of chemical-dependent protein switches for controlling intracellular electron transfer
Abstract
One concern with controlling electron flow in cells is the lack of biomolecules that directly couple the sensing of environmental conditions to electron transfer efficiency. To overcome this protein component limitation, we randomly inserted the ligand binding domain (LBD) from the human estrogen receptor (ER) into a thermostable 2Fe-2S ferredoxin (Fd) from Mastigocladus laminosus and used a bacterial selection to identify Fd-LBD fusion proteins that support electron transfer from a Fd-NADP reductase (FNR) to a Fd-dependent sulfite reductase (SIR). Mapping LBD insertion sites onto structure revealed that Fd tolerates domain insertion adjacent to or within the tetracysteine motif that coordinates the 2Fe-2S metallocluster. With both classes of the fusion proteins, cellular ET was enhanced by the ER antagonist 4-hydroxytamoxifen, albeit to different extents. In addition, one of Fds arising from ER-LBD insertion within the tetracysteine motif was found to acquire an oxygen-tolerant 2Fe-2S cluster, suggesting that ET is regulated through post-translational ligand binding. Furthermore, these findings show that domain insertion can be used to create protein electron carriers whose ET is regulated by environmental conditions. These metalloprotein switches are expected to be useful for achieving fast regulation of ET in engineered metabolic pathways and between electroactive bacteria and conductive materials.
- Authors:
-
- Rice Univ., Houston, TX (United States)
- Publication Date:
- Research Org.:
- Rice Univ., Houston, TX (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES); US Department of the Navy, Office of Naval Research (ONR)
- OSTI Identifier:
- 1594060
- Alternate Identifier(s):
- OSTI ID: 1570514
- Grant/Contract Number:
- SC0014462; N00014‐17‐1‐2639; DE‐SC0014462
- Resource Type:
- Accepted Manuscript
- Journal Name:
- AIChE Journal
- Additional Journal Information:
- Journal Volume: 66; Journal Issue: 3; Journal ID: ISSN 0001-1541
- Publisher:
- American Institute of Chemical Engineers
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 36 MATERIALS SCIENCE; metalloprotein; ferredoxin; electron transfer; microbiology; growth selection; allostery; domain insertion
Citation Formats
Wu, Bingyan, Atkinson, Joshua T., Kahanda, Dimithree, Bennett, George N., and Silberg, Jonathan J. Combinatorial design of chemical-dependent protein switches for controlling intracellular electron transfer. United States: N. p., 2019.
Web. doi:10.1002/aic.16796.
Wu, Bingyan, Atkinson, Joshua T., Kahanda, Dimithree, Bennett, George N., & Silberg, Jonathan J. Combinatorial design of chemical-dependent protein switches for controlling intracellular electron transfer. United States. doi:10.1002/aic.16796.
Wu, Bingyan, Atkinson, Joshua T., Kahanda, Dimithree, Bennett, George N., and Silberg, Jonathan J. Tue .
"Combinatorial design of chemical-dependent protein switches for controlling intracellular electron transfer". United States. doi:10.1002/aic.16796. https://www.osti.gov/servlets/purl/1594060.
@article{osti_1594060,
title = {Combinatorial design of chemical-dependent protein switches for controlling intracellular electron transfer},
author = {Wu, Bingyan and Atkinson, Joshua T. and Kahanda, Dimithree and Bennett, George N. and Silberg, Jonathan J.},
abstractNote = {One concern with controlling electron flow in cells is the lack of biomolecules that directly couple the sensing of environmental conditions to electron transfer efficiency. To overcome this protein component limitation, we randomly inserted the ligand binding domain (LBD) from the human estrogen receptor (ER) into a thermostable 2Fe-2S ferredoxin (Fd) from Mastigocladus laminosus and used a bacterial selection to identify Fd-LBD fusion proteins that support electron transfer from a Fd-NADP reductase (FNR) to a Fd-dependent sulfite reductase (SIR). Mapping LBD insertion sites onto structure revealed that Fd tolerates domain insertion adjacent to or within the tetracysteine motif that coordinates the 2Fe-2S metallocluster. With both classes of the fusion proteins, cellular ET was enhanced by the ER antagonist 4-hydroxytamoxifen, albeit to different extents. In addition, one of Fds arising from ER-LBD insertion within the tetracysteine motif was found to acquire an oxygen-tolerant 2Fe-2S cluster, suggesting that ET is regulated through post-translational ligand binding. Furthermore, these findings show that domain insertion can be used to create protein electron carriers whose ET is regulated by environmental conditions. These metalloprotein switches are expected to be useful for achieving fast regulation of ET in engineered metabolic pathways and between electroactive bacteria and conductive materials.},
doi = {10.1002/aic.16796},
journal = {AIChE Journal},
number = 3,
volume = 66,
place = {United States},
year = {2019},
month = {9}
}
Web of Science
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