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Expression and purification of the mammalian translocator protein for structural studies

The translocator protein (TSPO) is an 18 kDa polytopic membrane protein of the outer mitochondrial membrane, abundantly present in the steroid-synthesising cells. TSPO has been linked to a number of disorders, and it is recognised as a promising drug target with a range of potential medical applicat...

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Detalles Bibliográficos
Autores principales: Graeber, Elisabeth, Korkhov, Volodymyr M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5999236/
https://www.ncbi.nlm.nih.gov/pubmed/29897975
http://dx.doi.org/10.1371/journal.pone.0198832
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author Graeber, Elisabeth
Korkhov, Volodymyr M.
author_facet Graeber, Elisabeth
Korkhov, Volodymyr M.
author_sort Graeber, Elisabeth
collection PubMed
description The translocator protein (TSPO) is an 18 kDa polytopic membrane protein of the outer mitochondrial membrane, abundantly present in the steroid-synthesising cells. TSPO has been linked to a number of disorders, and it is recognised as a promising drug target with a range of potential medical applications. Structural and biochemical characterisation of a mammalian TSPO requires expression and purification of the protein of high quality in sufficiently large quantities. Here we describe detailed procedures for heterologous expression and purification of mammalian TSPO in HEK293 cells. We demonstrate that the established procedures can be used for untagged TSPO as well as for C-terminally fused TSPO constructs. Our protocol can be routinely used to generate high-quality TSPO preparations for biochemical and structural studies.
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spelling pubmed-59992362018-06-21 Expression and purification of the mammalian translocator protein for structural studies Graeber, Elisabeth Korkhov, Volodymyr M. PLoS One Research Article The translocator protein (TSPO) is an 18 kDa polytopic membrane protein of the outer mitochondrial membrane, abundantly present in the steroid-synthesising cells. TSPO has been linked to a number of disorders, and it is recognised as a promising drug target with a range of potential medical applications. Structural and biochemical characterisation of a mammalian TSPO requires expression and purification of the protein of high quality in sufficiently large quantities. Here we describe detailed procedures for heterologous expression and purification of mammalian TSPO in HEK293 cells. We demonstrate that the established procedures can be used for untagged TSPO as well as for C-terminally fused TSPO constructs. Our protocol can be routinely used to generate high-quality TSPO preparations for biochemical and structural studies. Public Library of Science 2018-06-13 /pmc/articles/PMC5999236/ /pubmed/29897975 http://dx.doi.org/10.1371/journal.pone.0198832 Text en © 2018 Graeber, Korkhov http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Graeber, Elisabeth
Korkhov, Volodymyr M.
Expression and purification of the mammalian translocator protein for structural studies
title Expression and purification of the mammalian translocator protein for structural studies
title_full Expression and purification of the mammalian translocator protein for structural studies
title_fullStr Expression and purification of the mammalian translocator protein for structural studies
title_full_unstemmed Expression and purification of the mammalian translocator protein for structural studies
title_short Expression and purification of the mammalian translocator protein for structural studies
title_sort expression and purification of the mammalian translocator protein for structural studies
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5999236/
https://www.ncbi.nlm.nih.gov/pubmed/29897975
http://dx.doi.org/10.1371/journal.pone.0198832
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