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

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.

doi:10.1021/jacs.5b11734

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

Journal Article · Thu Dec 24 00:00:00 EST 2015 · Journal of the American Chemical Society

DOI:https://doi.org/10.1021/jacs.5b11734· OSTI ID:1713208

Aussignargues, Clément ^[1]; Pandelia, Maria-Eirini ^[2]; Sutter, Markus ^[3]; Plegaria, Jefferson S. ^[4]; Zarzycki, Jan ^[4]; Turmo, Aiko ^[4]; Huang, Jingcheng ^[4]; Ducat, Daniel C. ^[4]; Hegg, Eric L. ^[5]; Gibney, Brian R. ^[6]; Kerfeld, Cheryl A. ^[7]

Michigan State Univ., East Lansing, MI (United States). MSU DOE Plant Research Lab.; MSU DOE Plant Research lab
Pennsylvania State Univ., University Park, PA (United States)
Michigan State Univ., East Lansing, MI (United States). MSU DOE Plant Research Lab.; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Michigan State Univ., East Lansing, MI (United States). MSU DOE Plant Research Lab.
Michigan State Univ., East Lansing, MI (United States)
Brooklyn College, Brooklyn, NY (United States)
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)

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.

View Accepted Manuscript (DOE)

Research Organization:: Michigan State Univ., East Lansing, MI (United States). MSU-DOE Plant Research Laboratory

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: FG02-91ER20021

OSTI ID:: 1713208

Journal Information:: Journal of the American Chemical Society, Journal Name: Journal of the American Chemical Society Journal Issue: 16 Vol. 138; ISSN 0002-7863

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

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