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Preparation of scFv stabilized chromatosomes for single-particle cryo-EM structure determination
The chromatosome, a nucleosome bound to a histone H1, is the structural unit of metazoan chromatin. Determination of the high-resolution structure of the chromatosome is challenging due to the dynamic nature of H1 binding. Here, we present a protocol for purifying an optimized single-chain antibody...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7994535/ https://www.ncbi.nlm.nih.gov/pubmed/33786462 http://dx.doi.org/10.1016/j.xpro.2021.100396 |
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author | Zhou, Bing-Rui Bai, Yawen |
author_facet | Zhou, Bing-Rui Bai, Yawen |
author_sort | Zhou, Bing-Rui |
collection | PubMed |
description | The chromatosome, a nucleosome bound to a histone H1, is the structural unit of metazoan chromatin. Determination of the high-resolution structure of the chromatosome is challenging due to the dynamic nature of H1 binding. Here, we present a protocol for purifying an optimized single-chain antibody variable fragment (scFv) that can be used to stabilize the chromatosome for single-particle cryo-EM studies. This protocol facilitates high-resolution cryo-EM structure determination of nucleosomes with a natural DNA sequence, chromatosomes, and other protein nucleosome complexes. For complete details on the use and execution of this protocol, please refer to Zhou et al. (2021). |
format | Online Article Text |
id | pubmed-7994535 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-79945352021-03-29 Preparation of scFv stabilized chromatosomes for single-particle cryo-EM structure determination Zhou, Bing-Rui Bai, Yawen STAR Protoc Protocol The chromatosome, a nucleosome bound to a histone H1, is the structural unit of metazoan chromatin. Determination of the high-resolution structure of the chromatosome is challenging due to the dynamic nature of H1 binding. Here, we present a protocol for purifying an optimized single-chain antibody variable fragment (scFv) that can be used to stabilize the chromatosome for single-particle cryo-EM studies. This protocol facilitates high-resolution cryo-EM structure determination of nucleosomes with a natural DNA sequence, chromatosomes, and other protein nucleosome complexes. For complete details on the use and execution of this protocol, please refer to Zhou et al. (2021). Elsevier 2021-03-20 /pmc/articles/PMC7994535/ /pubmed/33786462 http://dx.doi.org/10.1016/j.xpro.2021.100396 Text en http://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 | Protocol Zhou, Bing-Rui Bai, Yawen Preparation of scFv stabilized chromatosomes for single-particle cryo-EM structure determination |
title | Preparation of scFv stabilized chromatosomes for single-particle cryo-EM structure determination |
title_full | Preparation of scFv stabilized chromatosomes for single-particle cryo-EM structure determination |
title_fullStr | Preparation of scFv stabilized chromatosomes for single-particle cryo-EM structure determination |
title_full_unstemmed | Preparation of scFv stabilized chromatosomes for single-particle cryo-EM structure determination |
title_short | Preparation of scFv stabilized chromatosomes for single-particle cryo-EM structure determination |
title_sort | preparation of scfv stabilized chromatosomes for single-particle cryo-em structure determination |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7994535/ https://www.ncbi.nlm.nih.gov/pubmed/33786462 http://dx.doi.org/10.1016/j.xpro.2021.100396 |
work_keys_str_mv | AT zhoubingrui preparationofscfvstabilizedchromatosomesforsingleparticlecryoemstructuredetermination AT baiyawen preparationofscfvstabilizedchromatosomesforsingleparticlecryoemstructuredetermination |