Cesium based phasing of macromolecules: a general easy to use approach for solving the phase problem

Abstract Over the last decades the phase problem in macromolecular x-ray crystallography has become more controllable as methods and approaches have diversified and improved. However, solving the phase problem is still one of the biggest obstacles on the way of successfully determining a crystal str...

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Autores principales:	Wolfgang Koelmel, Jochen Kuper, Caroline Kisker
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4567078fe1304adea982cfe5ed2508da
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spelling	oai:doaj.org-article:4567078fe1304adea982cfe5ed2508da2021-12-02T18:53:14ZCesium based phasing of macromolecules: a general easy to use approach for solving the phase problem10.1038/s41598-021-95186-12045-2322https://doaj.org/article/4567078fe1304adea982cfe5ed2508da2021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-95186-1https://doaj.org/toc/2045-2322Abstract Over the last decades the phase problem in macromolecular x-ray crystallography has become more controllable as methods and approaches have diversified and improved. However, solving the phase problem is still one of the biggest obstacles on the way of successfully determining a crystal structure. To overcome this caveat, we have utilized the anomalous scattering properties of the heavy alkali metal cesium. We investigated the introduction of cesium in form of cesium chloride during the three major steps of protein treatment in crystallography: purification, crystallization, and cryo-protection. We derived a step-wise procedure encompassing a “quick-soak”-only approach and a combined approach of CsCl supplement during purification and cryo-protection. This procedure was successfully applied on two different proteins: (i) Lysozyme and (ii) as a proof of principle, a construct consisting of the PH domain of the TFIIH subunit p62 from Chaetomium thermophilum for de novo structure determination. Usage of CsCl thus provides a versatile, general, easy to use, and low cost phasing strategy.Wolfgang KoelmelJochen KuperCaroline KiskerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Wolfgang Koelmel Jochen Kuper Caroline Kisker Cesium based phasing of macromolecules: a general easy to use approach for solving the phase problem
description	Abstract Over the last decades the phase problem in macromolecular x-ray crystallography has become more controllable as methods and approaches have diversified and improved. However, solving the phase problem is still one of the biggest obstacles on the way of successfully determining a crystal structure. To overcome this caveat, we have utilized the anomalous scattering properties of the heavy alkali metal cesium. We investigated the introduction of cesium in form of cesium chloride during the three major steps of protein treatment in crystallography: purification, crystallization, and cryo-protection. We derived a step-wise procedure encompassing a “quick-soak”-only approach and a combined approach of CsCl supplement during purification and cryo-protection. This procedure was successfully applied on two different proteins: (i) Lysozyme and (ii) as a proof of principle, a construct consisting of the PH domain of the TFIIH subunit p62 from Chaetomium thermophilum for de novo structure determination. Usage of CsCl thus provides a versatile, general, easy to use, and low cost phasing strategy.
format	article
author	Wolfgang Koelmel Jochen Kuper Caroline Kisker
author_facet	Wolfgang Koelmel Jochen Kuper Caroline Kisker
author_sort	Wolfgang Koelmel
title	Cesium based phasing of macromolecules: a general easy to use approach for solving the phase problem
title_short	Cesium based phasing of macromolecules: a general easy to use approach for solving the phase problem
title_full	Cesium based phasing of macromolecules: a general easy to use approach for solving the phase problem
title_fullStr	Cesium based phasing of macromolecules: a general easy to use approach for solving the phase problem
title_full_unstemmed	Cesium based phasing of macromolecules: a general easy to use approach for solving the phase problem
title_sort	cesium based phasing of macromolecules: a general easy to use approach for solving the phase problem
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/4567078fe1304adea982cfe5ed2508da
work_keys_str_mv	AT wolfgangkoelmel cesiumbasedphasingofmacromoleculesageneraleasytouseapproachforsolvingthephaseproblem AT jochenkuper cesiumbasedphasingofmacromoleculesageneraleasytouseapproachforsolvingthephaseproblem AT carolinekisker cesiumbasedphasingofmacromoleculesageneraleasytouseapproachforsolvingthephaseproblem
_version_	1718377322984243200

Cesium based phasing of macromolecules: a general easy to use approach for solving the phase problem

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