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Title: Structure and Function of a Bacterial Microcompartment Shell Protein Engineered to Bind a [4Fe-4S] Cluster

Abstract

Bacterial microcompartments (BMCs) are self-assembling organelles composed of a selectively permeable protein shell and encapsulated enzymes. They are considered promising templates for the engineering of designed bionanoreactors for biotechnology. In particular, encapsulation of oxidoreductive reactions requiring electron transfer between the lumen of the BMC and the cytosol relies on the ability to conduct electrons across the shell. We determined the crystal structure of a component protein of a synthetic BMC shell, which informed the rational design of a [4Fe-4S] cluster-binding site in its pore. Here, we also solved the structure of the [4Fe-4S] cluster-bound, engineered protein to 1.8 Å resolution, providing the first structure of a BMC shell protein containing a metal center. The [4Fe-4S] cluster was characterized by optical and EPR spectroscopies; it has a reduction potential of -370 mV vs the standard hydrogen electrode (SHE) and is stable through redox cycling. This remarkable stability may be attributable to the hydrogen-bonding network provided by the main chain of the protein scaffold. The properties of the [4Fe-4S] cluster resemble those in low-potential bacterial ferredoxins, while its ligation to three cysteine residues is reminiscent of enzymes such as aconitase and radical S-adenosymethionine (SAM) enzymes. This engineered shell protein provides the foundationmore » for conferring electron-transfer functionality to BMC shells.« less

Authors:
 [1];  [2];  [3];  [1];  [1];  [1];  [1];  [1];  [4];  [5];  [6]
  1. Michigan State Univ., East Lansing, MI (United States). MSU DOE Plant Research Lab.
  2. Pennsylvania State Univ., University Park, PA (United States)
  3. Michigan State Univ., East Lansing, MI (United States). MSU DOE Plant Research Lab.; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  4. Michigan State Univ., East Lansing, MI (United States)
  5. Brooklyn College, Brooklyn, NY (United States)
  6. Michigan State Univ., East Lansing, MI (United States). MSU DOE Plant Research Lab.; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Berkeley, CA (United States); Berkeley Synthetic Biology Inst., Berkeley, CA (United States)
Publication Date:
Research Org.:
Michigan State Univ., East Lansing, MI (United States). MSU DOE Plant Research Lab.
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1713208
Grant/Contract Number:  
FG02-91ER20021
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Journal of the American Chemical Society
Additional Journal Information:
Journal Volume: 138; Journal Issue: 16; Journal ID: ISSN 0002-7863
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Aussignargues, Clément, Pandelia, Maria-Eirini, Sutter, Markus, Plegaria, Jefferson S., Zarzycki, Jan, Turmo, Aiko, Huang, Jingcheng, Ducat, Daniel C., Hegg, Eric L., Gibney, Brian R., and Kerfeld, Cheryl A. Structure and Function of a Bacterial Microcompartment Shell Protein Engineered to Bind a [4Fe-4S] Cluster. United States: N. p., 2015. Web. doi:10.1021/jacs.5b11734.
Aussignargues, Clément, Pandelia, Maria-Eirini, Sutter, Markus, Plegaria, Jefferson S., Zarzycki, Jan, Turmo, Aiko, Huang, Jingcheng, Ducat, Daniel C., Hegg, Eric L., Gibney, Brian R., & Kerfeld, Cheryl A. Structure and Function of a Bacterial Microcompartment Shell Protein Engineered to Bind a [4Fe-4S] Cluster. United States. https://doi.org/10.1021/jacs.5b11734
Aussignargues, Clément, Pandelia, Maria-Eirini, Sutter, Markus, Plegaria, Jefferson S., Zarzycki, Jan, Turmo, Aiko, Huang, Jingcheng, Ducat, Daniel C., Hegg, Eric L., Gibney, Brian R., and Kerfeld, Cheryl A. Thu . "Structure and Function of a Bacterial Microcompartment Shell Protein Engineered to Bind a [4Fe-4S] Cluster". United States. https://doi.org/10.1021/jacs.5b11734. https://www.osti.gov/servlets/purl/1713208.
@article{osti_1713208,
title = {Structure and Function of a Bacterial Microcompartment Shell Protein Engineered to Bind a [4Fe-4S] Cluster},
author = {Aussignargues, Clément and Pandelia, Maria-Eirini and Sutter, Markus and Plegaria, Jefferson S. and Zarzycki, Jan and Turmo, Aiko and Huang, Jingcheng and Ducat, Daniel C. and Hegg, Eric L. and Gibney, Brian R. and Kerfeld, Cheryl A.},
abstractNote = {Bacterial microcompartments (BMCs) are self-assembling organelles composed of a selectively permeable protein shell and encapsulated enzymes. They are considered promising templates for the engineering of designed bionanoreactors for biotechnology. In particular, encapsulation of oxidoreductive reactions requiring electron transfer between the lumen of the BMC and the cytosol relies on the ability to conduct electrons across the shell. We determined the crystal structure of a component protein of a synthetic BMC shell, which informed the rational design of a [4Fe-4S] cluster-binding site in its pore. Here, we also solved the structure of the [4Fe-4S] cluster-bound, engineered protein to 1.8 Å resolution, providing the first structure of a BMC shell protein containing a metal center. The [4Fe-4S] cluster was characterized by optical and EPR spectroscopies; it has a reduction potential of -370 mV vs the standard hydrogen electrode (SHE) and is stable through redox cycling. This remarkable stability may be attributable to the hydrogen-bonding network provided by the main chain of the protein scaffold. The properties of the [4Fe-4S] cluster resemble those in low-potential bacterial ferredoxins, while its ligation to three cysteine residues is reminiscent of enzymes such as aconitase and radical S-adenosymethionine (SAM) enzymes. This engineered shell protein provides the foundation for conferring electron-transfer functionality to BMC shells.},
doi = {10.1021/jacs.5b11734},
url = {https://www.osti.gov/biblio/1713208}, journal = {Journal of the American Chemical Society},
issn = {0002-7863},
number = 16,
volume = 138,
place = {United States},
year = {2015},
month = {12}
}

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