TY - JOUR
T1 - Complete and cooperative in vitro assembly of computationally designed self-assembling protein nanomaterials
AU - Wargacki, Adam J
AU - Wörner, Tobias P
AU - van de Waterbeemd, Michiel
AU - Ellis, Daniel
AU - Heck, Albert J R
AU - King, Neil P
N1 - Funding Information:
We thank Rashmi Ravichandran and the Nanoparticle Core Laboratory at the Institute for Protein Design for protein production and purification, Brooke Fiala for electron microscopy of in vitro-assembled I53-40 and I53-50, and members of the King laboratory for comments on the manuscript. T.P.W., M.V.D.W., and A.J.R.H. acknowledge support from the Netherlands Organization for Scientific Research (NWO) funding the Netherlands Proteomics Centre through the X-omics Road Map program (project 184.034.019) and the EU Horizon 2020 program INFRAIA project Epic-XS (Project 823839). A.W., D.R.E., and N.P.K. gratefully acknowledge support from the National Science Foundation (NSF CHE 1629214), DARPA (W911NF-17-2-0020), and the Bill & Melinda Gates Foundation (OPP1156262).
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/2/9
Y1 - 2021/2/9
N2 - Recent advances in computational methods have enabled the predictive design of self-assembling protein nanomaterials with atomic-level accuracy. These design strategies focus exclusively on a single target structure, without consideration of the mechanism or dynamics of assembly. However, understanding the assembly process, and in particular its robustness to perturbation, will be critical for translating this class of materials into useful technologies. Here we investigate the assembly of two computationally designed, 120-subunit icosahedral complexes in detail using several complementary biochemical methods. We found that assembly of each material from its two constituent protein building blocks was highly cooperative and yielded exclusively complete, 120-subunit complexes except in one non-stoichiometric regime for one of the materials. Our results suggest that in vitro assembly provides a robust and controllable route for the manufacture of designed protein nanomaterials and confirm that cooperative assembly can be an intrinsic, rather than evolved, feature of hierarchically structured protein complexes.
AB - Recent advances in computational methods have enabled the predictive design of self-assembling protein nanomaterials with atomic-level accuracy. These design strategies focus exclusively on a single target structure, without consideration of the mechanism or dynamics of assembly. However, understanding the assembly process, and in particular its robustness to perturbation, will be critical for translating this class of materials into useful technologies. Here we investigate the assembly of two computationally designed, 120-subunit icosahedral complexes in detail using several complementary biochemical methods. We found that assembly of each material from its two constituent protein building blocks was highly cooperative and yielded exclusively complete, 120-subunit complexes except in one non-stoichiometric regime for one of the materials. Our results suggest that in vitro assembly provides a robust and controllable route for the manufacture of designed protein nanomaterials and confirm that cooperative assembly can be an intrinsic, rather than evolved, feature of hierarchically structured protein complexes.
UR - http://www.scopus.com/inward/record.url?scp=85100823285&partnerID=8YFLogxK
U2 - 10.1038/s41467-021-21251-y
DO - 10.1038/s41467-021-21251-y
M3 - Article
C2 - 33563988
SN - 2041-1723
VL - 12
SP - 1
EP - 14
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 883
ER -