Pore structure controls stability and molecular flux in engineered protein cages-Reference-Cited by-同舟云学术

Pore structure controls stability and molecular flux in engineered protein cages

Published:2022-02-04 Issue:5 Volume:8 Page:
ISSN:2375-2548
Container-title:Science Advances
language:en
Short-container-title:Sci. Adv.

Author:

Adamson Lachlan S. R.¹²^ORCID,Tasneem Nuren¹^ORCID,Andreas Michael P.³⁴^ORCID,Close William⁵^ORCID,Jenner Eric N.¹^ORCID,Szyszka Taylor N.¹^ORCID,Young Reginald¹,Cheah Li Chen²⁶^ORCID,Norman Alexander¹⁷,MacDermott-Opeskin Hugo I.⁸^ORCID,O’Mara Megan L.⁸^ORCID,Sainsbury Frank²⁶⁹^ORCID,Giessen Tobias W.³⁴^ORCID,Lau Yu Heng¹⁷¹⁰^ORCID

Affiliation:

1. School of Chemistry, The University of Sydney, Camperdown, NSW 2006, Australia.

2. CSIRO Future Science Platform in Synthetic Biology, Commonwealth Scientific and Industrial Research Organisation (CSIRO), 41 Boggo Road, Dutton Park, QLD 4102, Australia.

3. Department of Biomedical Engineering, University of Michigan Medical School, Ann Arbor, MI, USA.

4. Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI, USA.

5. Australian Centre for Microscopy and Microanalysis, The University of Sydney, Camperdown, NSW 2006, Australia.

6. Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia, QLD 4072, Australia.

7. Australian Research Council Centre of Excellence for Innovations in Peptide and Protein Science, The University of Sydney, Camperdown, NSW 2006, Australia.

8. Research School of Chemistry, The Australian National University, Canberra, ACT 2601, Australia.

9. Centre for Cell Factories and Biopolymers, Griffith Institute for Drug Discovery, Griffith University, Nathan, QLD 4111, Australia.

10. The University of Sydney Nano Institute, The University of Sydney, Campderdown, NSW 2006, Australia.

Abstract

Protein cages are a common architectural motif used by living organisms to compartmentalize and control biochemical reactions. While engineered protein cages have featured in the construction of nanoreactors and synthetic organelles, relatively little is known about the underlying molecular parameters that govern stability and flux through their pores. In this work, we systematically designed 24 variants of the Thermotoga maritima encapsulin cage, featuring pores of different sizes and charges. Twelve pore variants were successfully assembled and purified, including eight designs with exceptional thermal stability. While negatively charged mutations were better tolerated, we were able to form stable assemblies covering a full range of pore sizes and charges, as observed in seven new cryo-EM structures at 2.5- to 3.6-Å resolution. Molecular dynamics simulations and stopped-flow experiments revealed the importance of considering both pore size and charge, together with flexibility and rate-determining steps, when designing protein cages for controlling molecular flux.

Publisher

American Association for the Advancement of Science (AAAS)

Subject

Multidisciplinary

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