The rapid “teabag” method for high-end purification of membrane proteins

Abstract Overproduction and purification of membrane proteins are generally challenging and time-consuming procedures due to low expression levels, misfolding, and low stability once extracted from the membrane. Reducing processing steps and shortening the timespan for purification represent attract...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Jenny Hering, Julie Winkel Missel, Liying Zhang, Anders Gunnarsson, Marie Castaldo, Per Amstrup Pedersen, Margareta Ek, Pontus Gourdon, Harm Jan Snijder
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4c535154a4b94033a5001d882e93a6d6
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:4c535154a4b94033a5001d882e93a6d6
record_format	dspace
spelling	oai:doaj.org-article:4c535154a4b94033a5001d882e93a6d62021-12-02T18:51:07ZThe rapid “teabag” method for high-end purification of membrane proteins10.1038/s41598-020-73285-92045-2322https://doaj.org/article/4c535154a4b94033a5001d882e93a6d62020-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-73285-9https://doaj.org/toc/2045-2322Abstract Overproduction and purification of membrane proteins are generally challenging and time-consuming procedures due to low expression levels, misfolding, and low stability once extracted from the membrane. Reducing processing steps and shortening the timespan for purification represent attractive approaches to overcome some of these challenges. We have therefore compared a fast “teabag” purification method with conventional purification for five different membrane proteins (MraY, AQP10, ClC-1, PAR2 and KCC2). Notably, this new approach reduces the purification time significantly, and the quality of the purified membrane proteins is equal to or exceeds conventional methods as assessed by size exclusion chromatography, SDS-PAGE and downstream applications such as ITC, crystallization and cryo-EM. Furthermore, the method is scalable, applicable to a range of affinity resins and allows for parallelization. Consequently, the technique has the potential to substantially simplify purification efforts of membrane proteins in basic and applied sciences.Jenny HeringJulie Winkel MisselLiying ZhangAnders GunnarssonMarie CastaldoPer Amstrup PedersenMargareta EkPontus GourdonHarm Jan SnijderNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-11 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jenny Hering Julie Winkel Missel Liying Zhang Anders Gunnarsson Marie Castaldo Per Amstrup Pedersen Margareta Ek Pontus Gourdon Harm Jan Snijder The rapid “teabag” method for high-end purification of membrane proteins
description	Abstract Overproduction and purification of membrane proteins are generally challenging and time-consuming procedures due to low expression levels, misfolding, and low stability once extracted from the membrane. Reducing processing steps and shortening the timespan for purification represent attractive approaches to overcome some of these challenges. We have therefore compared a fast “teabag” purification method with conventional purification for five different membrane proteins (MraY, AQP10, ClC-1, PAR2 and KCC2). Notably, this new approach reduces the purification time significantly, and the quality of the purified membrane proteins is equal to or exceeds conventional methods as assessed by size exclusion chromatography, SDS-PAGE and downstream applications such as ITC, crystallization and cryo-EM. Furthermore, the method is scalable, applicable to a range of affinity resins and allows for parallelization. Consequently, the technique has the potential to substantially simplify purification efforts of membrane proteins in basic and applied sciences.
format	article
author	Jenny Hering Julie Winkel Missel Liying Zhang Anders Gunnarsson Marie Castaldo Per Amstrup Pedersen Margareta Ek Pontus Gourdon Harm Jan Snijder
author_facet	Jenny Hering Julie Winkel Missel Liying Zhang Anders Gunnarsson Marie Castaldo Per Amstrup Pedersen Margareta Ek Pontus Gourdon Harm Jan Snijder
author_sort	Jenny Hering
title	The rapid “teabag” method for high-end purification of membrane proteins
title_short	The rapid “teabag” method for high-end purification of membrane proteins
title_full	The rapid “teabag” method for high-end purification of membrane proteins
title_fullStr	The rapid “teabag” method for high-end purification of membrane proteins
title_full_unstemmed	The rapid “teabag” method for high-end purification of membrane proteins
title_sort	rapid “teabag” method for high-end purification of membrane proteins
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/4c535154a4b94033a5001d882e93a6d6
work_keys_str_mv	AT jennyhering therapidteabagmethodforhighendpurificationofmembraneproteins AT juliewinkelmissel therapidteabagmethodforhighendpurificationofmembraneproteins AT liyingzhang therapidteabagmethodforhighendpurificationofmembraneproteins AT andersgunnarsson therapidteabagmethodforhighendpurificationofmembraneproteins AT mariecastaldo therapidteabagmethodforhighendpurificationofmembraneproteins AT peramstruppedersen therapidteabagmethodforhighendpurificationofmembraneproteins AT margaretaek therapidteabagmethodforhighendpurificationofmembraneproteins AT pontusgourdon therapidteabagmethodforhighendpurificationofmembraneproteins AT harmjansnijder therapidteabagmethodforhighendpurificationofmembraneproteins AT jennyhering rapidteabagmethodforhighendpurificationofmembraneproteins AT juliewinkelmissel rapidteabagmethodforhighendpurificationofmembraneproteins AT liyingzhang rapidteabagmethodforhighendpurificationofmembraneproteins AT andersgunnarsson rapidteabagmethodforhighendpurificationofmembraneproteins AT mariecastaldo rapidteabagmethodforhighendpurificationofmembraneproteins AT peramstruppedersen rapidteabagmethodforhighendpurificationofmembraneproteins AT margaretaek rapidteabagmethodforhighendpurificationofmembraneproteins AT pontusgourdon rapidteabagmethodforhighendpurificationofmembraneproteins AT harmjansnijder rapidteabagmethodforhighendpurificationofmembraneproteins
_version_	1718377444923146240

The rapid “teabag” method for high-end purification of membrane proteins

Ejemplares similares