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De novo targeting to the cytoplasmic and luminal side of bacterial microcompartments

Lee, Matthew J., Mantell, Judith, Brown, Ian R., Fletcher, Jordan M., Verkade, Paul, Pickersgill, Richard W., Woolfson, Derek N., Frank, Stefanie, Warren, Martin J. (2018) De novo targeting to the cytoplasmic and luminal side of bacterial microcompartments. Nature Communications, 9 (1). p. 3413. ISSN 2041-1723. E-ISSN 2041-1723. (doi:10.1038/s41467-018-05922-x) (KAR id:68857)

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https://doi.org/10.1038/s41467-018-05922-x

Abstract

Bacterial microcompartments, BMCs, are proteinaceous organelles that encase a specific metabolic pathway within a semi-permeable protein shell. Short encapsulation peptides can direct cargo proteins to the lumen of the compartments. However, the fusion of such peptides to non-native proteins does not guarantee encapsulation and often causes aggregation. Here, we report an approach for targeting recombinant proteins to BMCs that utilizes specific de novo coiled-coil protein–protein interactions. Attachment of one coiled-coil module to PduA (a component of the BMC shell) allows targeting of a fluorescent protein fused to a cognate coiled-coil partner. This interaction takes place on the outer surface of the BMC. The redesign of PduA to generate an N-terminus on the luminal side of the BMC results in intact compartments to which proteins can still be targeted via the designed coiled-coil system. This study provides a strategy to display proteins on the surface or within the lumen of the BMCs.

Item Type: Article
DOI/Identification number: 10.1038/s41467-018-05922-x
Uncontrolled keywords: Metabolic engineering, Microbiology techniques, Protein design
Subjects: Q Science
Q Science > QR Microbiology
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: Martin Warren
Date Deposited: 30 Aug 2018 10:14 UTC
Last Modified: 29 May 2019 21:04 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/68857 (The current URI for this page, for reference purposes)
Verkade, Paul: https://orcid.org/0000-0002-2497-1026
Woolfson, Derek N.: https://orcid.org/0000-0002-0394-3202
Warren, Martin J.: https://orcid.org/0000-0002-6028-6456
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