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Title: Programmed loading and rapid purification of engineered bacterial microcompartment shells

Journal Article · · Nature Communications
 [1]; ORCiD logo [2];  [1]; ORCiD logo [2]
  1. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  2. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Michigan State Univ., East Lansing, MI (United States)

Bacterial microcompartments (BMCs) are selectively permeable proteinaceous organelles which encapsulate segments of metabolic pathways across bacterial phyla. They consist of an enzymatic core surrounded by a protein shell composed of multiple distinct proteins. Despite great potential in varied biotechnological applications, engineering efforts have been stymied by difficulties in their isolation and characterization and a dearth of robust methods for programming cores and shell permeability. We address these challenges by functionalizing shell proteins with affinity handles, enabling facile complementation-based affinity purification (CAP) and specific cargo docking sites for efficient encapsulation via covalent-linkage (EnCo). These shell functionalizations extend our knowledge of BMC architectural principles and enable the development of minimal shell systems of precisely defined structure and composition. The generalizability of CAP and EnCo will enable their application to functionally diverse microcompartment systems to facilitate both characterization of natural functions and the development of bespoke shells for selectively compartmentalizing proteins.

Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); Michigan State Univ., East Lansing, MI (United States). DOE Plant Research Laboratory
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
Grant/Contract Number:
AC02-05CH11231; FG02-91ER20021
OSTI ID:
1460586
Alternate ID(s):
OSTI ID: 1478349; OSTI ID: 1603659
Journal Information:
Nature Communications, Vol. 9, Issue 1; ISSN 2041-1723
Publisher:
Nature Publishing GroupCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 70 works
Citation information provided by
Web of Science

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Bacterial microcompartments: catalysis-enhancing metabolic modules for next generation metabolic and biomedical engineering journal October 2019
Spy&Go purification of SpyTag-proteins using pseudo-SpyCatcher to access an oligomerization toolbox journal April 2019
Encapsulation mechanisms and structural studies of GRM2 bacterial microcompartment particles journal January 2020
Covalent Functionalization of Bioengineered Polyhydroxyalkanoate Spheres Directed by Specific Protein-Protein Interactions journal February 2020
Single-Organelle Quantification Reveals Stoichiometric and Structural Variability of Carboxysomes Dependent on the Environment journal May 2019
Self‐Assembly Toolbox of Tailored Supramolecular Architectures Based on an Amphiphilic Protein Library journal June 2019
Engineering the PduT shell protein to modify the permeability of the 1,2-propanediol microcompartment of Salmonella journal December 2019
Encapsulation mechanisms and structural studies of GRM2 bacterial microcompartment particles journal January 2020
The Phenix software for automated determination of macromolecular structures journal September 2011
Functionalization of Bacterial Microcompartment Shell Proteins With Covalently Attached Heme journal January 2020
Programmable assembly of 2D crystalline protein arrays into covalently stacked 3D bionanomaterials journal January 2020
Investigating the functional and structural modulation of carboxysomes in Synechococcus elongatus PCC7942 text January 2019
Functionalization of Bacterial Microcompartment Shell Proteins With Covalently Attached Heme journal January 2020
Covalent Functionalization of Bioengineered Polyhydroxyalkanoate Spheres Directed by Specific Protein-Protein Interactions journal February 2020