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Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins
Significant advances in the past decade have enabled high-resolution structure determination of a vast variety of proteins by cryogenic electron microscopy single particle analysis. Despite improved sample preparation, next-generation imaging hardware, and advanced single particle analysis algorithm...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9562432/ https://www.ncbi.nlm.nih.gov/pubmed/36248264 http://dx.doi.org/10.1016/j.crstbi.2022.09.005 |
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author | Wentinck, Koen Gogou, Christos Meijer, Dimphna H. |
author_facet | Wentinck, Koen Gogou, Christos Meijer, Dimphna H. |
author_sort | Wentinck, Koen |
collection | PubMed |
description | Significant advances in the past decade have enabled high-resolution structure determination of a vast variety of proteins by cryogenic electron microscopy single particle analysis. Despite improved sample preparation, next-generation imaging hardware, and advanced single particle analysis algorithms, small proteins remain elusive for reconstruction due to low signal-to-noise and lack of distinctive structural features. Multiple efforts have therefore been directed at the development of size-increase techniques for small proteins. Here we review the latest methods for increasing effective molecular weight of proteins <100 kDa through target protein binding or target protein fusion - specifically by using nanobody-based assemblies, fusion tags, and symmetric scaffolds. Finally, we summarize these state-of-the-art techniques into a decision-tree to facilitate the design of tailored future approaches, and thus for further exploration of ever-smaller proteins that make up the largest part of the human genome. |
format | Online Article Text |
id | pubmed-9562432 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-95624322022-10-15 Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins Wentinck, Koen Gogou, Christos Meijer, Dimphna H. Curr Res Struct Biol Graphical Review Significant advances in the past decade have enabled high-resolution structure determination of a vast variety of proteins by cryogenic electron microscopy single particle analysis. Despite improved sample preparation, next-generation imaging hardware, and advanced single particle analysis algorithms, small proteins remain elusive for reconstruction due to low signal-to-noise and lack of distinctive structural features. Multiple efforts have therefore been directed at the development of size-increase techniques for small proteins. Here we review the latest methods for increasing effective molecular weight of proteins <100 kDa through target protein binding or target protein fusion - specifically by using nanobody-based assemblies, fusion tags, and symmetric scaffolds. Finally, we summarize these state-of-the-art techniques into a decision-tree to facilitate the design of tailored future approaches, and thus for further exploration of ever-smaller proteins that make up the largest part of the human genome. Elsevier 2022-10-02 /pmc/articles/PMC9562432/ /pubmed/36248264 http://dx.doi.org/10.1016/j.crstbi.2022.09.005 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Graphical Review Wentinck, Koen Gogou, Christos Meijer, Dimphna H. Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins |
title | Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins |
title_full | Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins |
title_fullStr | Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins |
title_full_unstemmed | Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins |
title_short | Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins |
title_sort | putting on molecular weight: enabling cryo-em structure determination of sub-100-kda proteins |
topic | Graphical Review |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9562432/ https://www.ncbi.nlm.nih.gov/pubmed/36248264 http://dx.doi.org/10.1016/j.crstbi.2022.09.005 |
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