Engineering and functionalization of large circular tandem repeat protein nanoparticles

Protein engineering has enabled the design of molecular scaffolds that display a wide variety of sizes, shapes, symmetries and subunit compositions. Symmetric protein-based nanoparticles that display multiple protein domains can exhibit enhanced functional properties due to increased avidity and imp...

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Bibliographic Details
Published inNature structural & molecular biology Vol. 27; no. 4; pp. 342 - 350
Main Authors Correnti, Colin E., Hallinan, Jazmine P., Doyle, Lindsey A., Ruff, Raymond O., Jaeger-Ruckstuhl, Carla A., Xu, Yuexin, Shen, Betty W., Qu, Amanda, Polkinghorn, Caley, Friend, Della J., Bandaranayake, Ashok D., Riddell, Stanley R., Kaiser, Brett K., Stoddard, Barry L., Bradley, Philip
Format Journal Article
LanguageEnglish
Published New York Nature Publishing Group US 01.04.2020
Nature Publishing Group
Subjects
101/28
13
13/31
631/250
631/45/612
82
82/103
82/80
82/83
Amino Acid Motifs - genetics
Avidity
Biochemistry
Biological Microscopy
Biomedical and Life Sciences
Cargo
Cell culture
Cell Culture Techniques
Cell surface
Circularity
Composition
Humans
Kinases
Life Sciences
Lymphocytes
Lymphocytes T
Membrane Biology
Methods
Models, Molecular
Molecular interactions
Nanoparticles
Nanoparticles - chemistry
Protein Domains - genetics
Protein Engineering
Protein research
Protein Stability
Protein Structure
Proteins
Proteins - chemistry
Proteins - genetics
Self-assembly
Structure
T cells
T-Lymphocytes - chemistry
Tandem Repeat Sequences - genetics
United States
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Summary:Protein engineering has enabled the design of molecular scaffolds that display a wide variety of sizes, shapes, symmetries and subunit compositions. Symmetric protein-based nanoparticles that display multiple protein domains can exhibit enhanced functional properties due to increased avidity and improved solution behavior and stability. Here we describe the creation and characterization of a computationally designed circular tandem repeat protein (cTRP) composed of 24 identical repeated motifs, which can display a variety of functional protein domains (cargo) at defined positions around its periphery. We demonstrate that cTRP nanoparticles can self-assemble from smaller individual subunits, can be produced from prokaryotic and human expression platforms, can employ a variety of cargo attachment strategies and can be used for applications (such as T-cell culture and expansion) requiring high-avidity molecular interactions on the cell surface. Circular nanoparticles self-assembled from designed tandem repeat proteins are functionalized by fusing different protein domains at defined positions and copy numbers. These constructs can activate T cells via high-avidity interactions on the cell surface.
Bibliography:P.B. conducted the computational protein design work, including fold design and identification of point mutations leading to alteration of self-assembly properties and generated the structural models used throughout the article. J.H., L.A.D., A.Q., C.P., B.K.K., B.L.S. and R.O.R. all conducted protein expression, purification and biochemical characterization experiments. B.W.S. conducted EM visualization studies. D.J.F. conducted surface plasmon resonance protein binding studies. Y.X., C.A.J., A.D.B. and S.R.R. designed and conducted T-cell staining studies. C.E.C. and B.L.S. designed functionalized cTRP constructs. C.E.C, B.K.K, B.L.S. and P.B. wrote the manuscript, which was edited extensively by all authors.
Author contributions
ISSN:1545-9993
1545-9985
DOI:10.1038/s41594-020-0397-5