A Suite of Engineered GFP Molecules for Oligomeric Scaffolding
Abstract
Applications ranging from synthetic biology to protein crystallization could be advanced by facile systems for connecting multiple proteins together in predefined spatial relationships. One approach to this goal is to engineer many distinct assembly forms of a single carrier protein or scaffold, to which other proteins of interest can then be readily attached. In this work we chose GFP as a scaffold and engineered many alternative oligomeric forms, driven by either specific disulfide bond formation or metal ion addition. We generated a wide range of spatial arrangements of GFP subunits from 11 different oligomeric variants, and determined their X-ray structures in a total of 33 distinct crystal forms. Furthermore, some of the oligomeric GFP variants show geometric polymorphism depending on conditions, while others show considerable geometric rigidity. Potential future applications of this system are discussed.
- Authors:
- Publication Date:
- Research Org.:
- Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER); USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1337421
- Alternate Identifier(s):
- OSTI ID: 1250980; OSTI ID: 1255154
- Report Number(s):
- LA-UR-15-29210
Journal ID: ISSN 0969-2126; S0969212615002890; PII: S0969212615002890
- Grant/Contract Number:
- FC02-02ER63421; AC02-06CH11357; P01 GM098177; T32GM007185; RR-15301; AC52-06NA25396
- Resource Type:
- Published Article
- Journal Name:
- Structure
- Additional Journal Information:
- Journal Name: Structure Journal Volume: 23 Journal Issue: 9; Journal ID: ISSN 0969-2126
- Publisher:
- Elsevier
- Country of Publication:
- United Kingdom
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; protein scaffolds; protein crystallization; protein engineering; protein assembly; synthetic biology
Citation Formats
Leibly, David J., Arbing, Mark A., Pashkov, Inna, DeVore, Natasha, Waldo, Geoffrey S., Terwilliger, Thomas C., and Yeates, Todd O. A Suite of Engineered GFP Molecules for Oligomeric Scaffolding. United Kingdom: N. p., 2015.
Web. doi:10.1016/j.str.2015.07.008.
Leibly, David J., Arbing, Mark A., Pashkov, Inna, DeVore, Natasha, Waldo, Geoffrey S., Terwilliger, Thomas C., & Yeates, Todd O. A Suite of Engineered GFP Molecules for Oligomeric Scaffolding. United Kingdom. https://doi.org/10.1016/j.str.2015.07.008
Leibly, David J., Arbing, Mark A., Pashkov, Inna, DeVore, Natasha, Waldo, Geoffrey S., Terwilliger, Thomas C., and Yeates, Todd O. Tue .
"A Suite of Engineered GFP Molecules for Oligomeric Scaffolding". United Kingdom. https://doi.org/10.1016/j.str.2015.07.008.
@article{osti_1337421,
title = {A Suite of Engineered GFP Molecules for Oligomeric Scaffolding},
author = {Leibly, David J. and Arbing, Mark A. and Pashkov, Inna and DeVore, Natasha and Waldo, Geoffrey S. and Terwilliger, Thomas C. and Yeates, Todd O.},
abstractNote = {Applications ranging from synthetic biology to protein crystallization could be advanced by facile systems for connecting multiple proteins together in predefined spatial relationships. One approach to this goal is to engineer many distinct assembly forms of a single carrier protein or scaffold, to which other proteins of interest can then be readily attached. In this work we chose GFP as a scaffold and engineered many alternative oligomeric forms, driven by either specific disulfide bond formation or metal ion addition. We generated a wide range of spatial arrangements of GFP subunits from 11 different oligomeric variants, and determined their X-ray structures in a total of 33 distinct crystal forms. Furthermore, some of the oligomeric GFP variants show geometric polymorphism depending on conditions, while others show considerable geometric rigidity. Potential future applications of this system are discussed.},
doi = {10.1016/j.str.2015.07.008},
journal = {Structure},
number = 9,
volume = 23,
place = {United Kingdom},
year = {Tue Sep 01 00:00:00 EDT 2015},
month = {Tue Sep 01 00:00:00 EDT 2015}
}
https://doi.org/10.1016/j.str.2015.07.008
Web of Science
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