A Suite of Engineered GFP Molecules for Oligomeric Scaffolding
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.
- Research Organization:
- Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)
- Sponsoring Organization:
- USDOE Office of Science (SC), Biological and Environmental Research (BER); USDOE Office of Science (SC), Basic Energy Sciences (BES)
- Grant/Contract Number:
- FC02-02ER63421; AC02-06CH11357; P01 GM098177; T32GM007185; RR-15301; AC52-06NA25396
- OSTI ID:
- 1337421
- Alternate ID(s):
- OSTI ID: 1250980; OSTI ID: 1255154
- Report Number(s):
- LA-UR-15-29210; S0969212615002890; PII: S0969212615002890
- Journal Information:
- Structure, Journal Name: Structure Vol. 23 Journal Issue: 9; ISSN 0969-2126
- Publisher:
- ElsevierCopyright Statement
- Country of Publication:
- United Kingdom
- Language:
- English
Web of Science
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