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:
-
- Michigan State Univ., East Lansing, MI (United States). MSU-DOE Plant Research Laboratory
- Mannheim Univ. of Applied Sciences (Germany)
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
- 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. doi: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 = {Wed Nov 20 00:00:00 EST 2019},
month = {Wed Nov 20 00:00:00 EST 2019}
}
Web of Science