Cargando…

Functionalization of the BCL6 BTB domain into a noncovalent crystallization chaperone

The production of diffraction-quality protein crystals is challenging and often requires bespoke, time-consuming and expensive strategies. A system has been developed in which the BCL6 BTB domain acts as a crystallization chaperone and promiscuous assembly block that may form the basis for affinity-...

Descripción completa

Detalles Bibliográficos
Autores principales: Zacharchenko, Thomas, Wright, Stephanie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7924223/
https://www.ncbi.nlm.nih.gov/pubmed/33708392
http://dx.doi.org/10.1107/S2052252520015754
_version_ 1783659045797756928
author Zacharchenko, Thomas
Wright, Stephanie
author_facet Zacharchenko, Thomas
Wright, Stephanie
author_sort Zacharchenko, Thomas
collection PubMed
description The production of diffraction-quality protein crystals is challenging and often requires bespoke, time-consuming and expensive strategies. A system has been developed in which the BCL6 BTB domain acts as a crystallization chaperone and promiscuous assembly block that may form the basis for affinity-capture crystallography. The protein of interest is expressed with a C-terminal tag that interacts with the BTB domain, and co-crystallization leads to its incorporation within a BTB-domain lattice. This strategy was used to solve the structure of the SH3 domain of human nebulin, a structure previously solved by NMR, at 1.56 Å resolution. This approach is simple and effective, requiring only routine protein complexation and crystallization screening, and should be applicable to a range of proteins.
format Online
Article
Text
id pubmed-7924223
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher International Union of Crystallography
record_format MEDLINE/PubMed
spelling pubmed-79242232021-03-10 Functionalization of the BCL6 BTB domain into a noncovalent crystallization chaperone Zacharchenko, Thomas Wright, Stephanie IUCrJ Research Letters The production of diffraction-quality protein crystals is challenging and often requires bespoke, time-consuming and expensive strategies. A system has been developed in which the BCL6 BTB domain acts as a crystallization chaperone and promiscuous assembly block that may form the basis for affinity-capture crystallography. The protein of interest is expressed with a C-terminal tag that interacts with the BTB domain, and co-crystallization leads to its incorporation within a BTB-domain lattice. This strategy was used to solve the structure of the SH3 domain of human nebulin, a structure previously solved by NMR, at 1.56 Å resolution. This approach is simple and effective, requiring only routine protein complexation and crystallization screening, and should be applicable to a range of proteins. International Union of Crystallography 2021-01-11 /pmc/articles/PMC7924223/ /pubmed/33708392 http://dx.doi.org/10.1107/S2052252520015754 Text en © Zacharchenko & Wright 2021 http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.http://creativecommons.org/licenses/by/4.0/
spellingShingle Research Letters
Zacharchenko, Thomas
Wright, Stephanie
Functionalization of the BCL6 BTB domain into a noncovalent crystallization chaperone
title Functionalization of the BCL6 BTB domain into a noncovalent crystallization chaperone
title_full Functionalization of the BCL6 BTB domain into a noncovalent crystallization chaperone
title_fullStr Functionalization of the BCL6 BTB domain into a noncovalent crystallization chaperone
title_full_unstemmed Functionalization of the BCL6 BTB domain into a noncovalent crystallization chaperone
title_short Functionalization of the BCL6 BTB domain into a noncovalent crystallization chaperone
title_sort functionalization of the bcl6 btb domain into a noncovalent crystallization chaperone
topic Research Letters
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7924223/
https://www.ncbi.nlm.nih.gov/pubmed/33708392
http://dx.doi.org/10.1107/S2052252520015754
work_keys_str_mv AT zacharchenkothomas functionalizationofthebcl6btbdomainintoanoncovalentcrystallizationchaperone
AT wrightstephanie functionalizationofthebcl6btbdomainintoanoncovalentcrystallizationchaperone