skip to main content
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Visualizing in Vivo Dynamics of Designer Nanoscaffolds

Abstract

Enzymes of natural biochemical pathways are routinely subcellularly organized in space and time in order to improve pathway efficacy and control. Designer scaffolding platforms are under development to confer similar benefits upon engineered pathways. Herein, we evaluate bacterial microcompartment shell (pfam0936-domain) proteins as modules for constructing well-defined nanometer scale scaffolds in vivo. We use a suite of visualization techniques to evaluate scaffold assembly and dynamics. We demonstrate recruitment of target cargo molecules onto assembled scaffolds by appending reciprocally interacting adaptor domains. These interactions can be refined by fine-tuning the scaffold expression level. Real-time observation of this system reveals a nucleation-limited step where multiple scaffolds initially form within a cell. Over time, nucleated scaffolds reorganize into a single intracellular assembly, likely due to interscaffold competition for protein subunits. Our results suggest design considerations for using self-assembling proteins as building blocks to construct nanoscaffolds, while also providing a platform to visualize scaffold-cargo dynamics in vivo.

Authors:
 [1];  [1];  [1];  [1];  [2];  [3]; ORCiD logo [4]; ORCiD logo [1]
  1. Michigan State Univ., East Lansing, MI (United States). MSU-DOE Plant Research Laboratory
  2. Mannheim Univ. of Applied Sciences (Germany)
  3. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  4. Michigan State Univ., East Lansing, MI (United States). MSU-DOE Plant Research Laboratory; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Publication Date:
Research Org.:
Michigan State Univ., East Lansing, MI (United States). MSU-DOE Plant Research Laboratory
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1608250
Alternate Identifier(s):
OSTI ID: 1735927
Grant/Contract Number:  
FG02-91ER20021
Resource Type:
Accepted Manuscript
Journal Name:
Nano Letters
Additional Journal Information:
Journal Volume: 20; Journal Issue: 1; Journal ID: ISSN 1530-6984
Publisher:
American Chemical Society
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Protein scaffold; self-assembly; nanotechnology; synthetic biology; bioengineering

Citation Formats

Young, Eric J., Sakkos, Jonathan K., Huang, Jingcheng, Wright, Jacob K., Kachel, Benjamin, Fuentes-Cabrera, Miguel, Kerfeld, Cheryl A., and Ducat, Daniel C.. Visualizing in Vivo Dynamics of Designer Nanoscaffolds. United States: N. p., 2019. Web. https://doi.org/10.1021/acs.nanolett.9b03651.
Young, Eric J., Sakkos, Jonathan K., Huang, Jingcheng, Wright, Jacob K., Kachel, Benjamin, Fuentes-Cabrera, Miguel, Kerfeld, Cheryl A., & Ducat, Daniel C.. Visualizing in Vivo Dynamics of Designer Nanoscaffolds. United States. https://doi.org/10.1021/acs.nanolett.9b03651
Young, Eric J., Sakkos, Jonathan K., Huang, Jingcheng, Wright, Jacob K., Kachel, Benjamin, Fuentes-Cabrera, Miguel, Kerfeld, Cheryl A., and Ducat, Daniel C.. Wed . "Visualizing in Vivo Dynamics of Designer Nanoscaffolds". United States. https://doi.org/10.1021/acs.nanolett.9b03651. https://www.osti.gov/servlets/purl/1608250.
@article{osti_1608250,
title = {Visualizing in Vivo Dynamics of Designer Nanoscaffolds},
author = {Young, Eric J. and Sakkos, Jonathan K. and Huang, Jingcheng and Wright, Jacob K. and Kachel, Benjamin and Fuentes-Cabrera, Miguel and Kerfeld, Cheryl A. and Ducat, Daniel C.},
abstractNote = {Enzymes of natural biochemical pathways are routinely subcellularly organized in space and time in order to improve pathway efficacy and control. Designer scaffolding platforms are under development to confer similar benefits upon engineered pathways. Herein, we evaluate bacterial microcompartment shell (pfam0936-domain) proteins as modules for constructing well-defined nanometer scale scaffolds in vivo. We use a suite of visualization techniques to evaluate scaffold assembly and dynamics. We demonstrate recruitment of target cargo molecules onto assembled scaffolds by appending reciprocally interacting adaptor domains. These interactions can be refined by fine-tuning the scaffold expression level. Real-time observation of this system reveals a nucleation-limited step where multiple scaffolds initially form within a cell. Over time, nucleated scaffolds reorganize into a single intracellular assembly, likely due to interscaffold competition for protein subunits. Our results suggest design considerations for using self-assembling proteins as building blocks to construct nanoscaffolds, while also providing a platform to visualize scaffold-cargo dynamics in vivo.},
doi = {10.1021/acs.nanolett.9b03651},
journal = {Nano Letters},
number = 1,
volume = 20,
place = {United States},
year = {2019},
month = {11}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Save / Share: