skip to main content

DOE PAGESDOE PAGES

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

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.
Authors:
 [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)
Publication Date:
Grant/Contract Number:
AC02-05CH11231
Type:
Published Article
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 9; Journal Issue: 1; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Research Org:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES
OSTI Identifier:
1460586
Alternate Identifier(s):
OSTI ID: 1478349

Hagen, Andrew, Sutter, Markus, Sloan, Nancy, and Kerfeld, Cheryl A. Programmed loading and rapid purification of engineered bacterial microcompartment shells. United States: N. p., Web. doi:10.1038/s41467-018-05162-z.
Hagen, Andrew, Sutter, Markus, Sloan, Nancy, & Kerfeld, Cheryl A. Programmed loading and rapid purification of engineered bacterial microcompartment shells. United States. doi:10.1038/s41467-018-05162-z.
Hagen, Andrew, Sutter, Markus, Sloan, Nancy, and Kerfeld, Cheryl A. 2018. "Programmed loading and rapid purification of engineered bacterial microcompartment shells". United States. doi:10.1038/s41467-018-05162-z.
@article{osti_1460586,
title = {Programmed loading and rapid purification of engineered bacterial microcompartment shells},
author = {Hagen, Andrew and Sutter, Markus and Sloan, Nancy and Kerfeld, Cheryl A.},
abstractNote = {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.},
doi = {10.1038/s41467-018-05162-z},
journal = {Nature Communications},
number = 1,
volume = 9,
place = {United States},
year = {2018},
month = {7}
}

Works referenced in this record:

Microcompartments for B12-Dependent 1,2-Propanediol Degradation Provide Protection from DNA and Cellular Damage by a Reactive Metabolic Intermediate
journal, February 2008
  • Sampson, E. M.; Bobik, T. A.
  • Journal of Bacteriology, Vol. 190, Issue 8, p. 2966-2971
  • DOI: 10.1128/JB.01925-07

Specific Covalent Labeling of Recombinant Protein Molecules Inside Live Cells
journal, July 1998