A phosphorylation tag for uranyl mediated protein purification and photo assisted tag removal.

Most protein purification procedures include an affinity tag fused to either the N or C-terminal end of the protein of interest as well as a procedure for tag removal. Tag removal is not straightforward and especially tag removal from the C-terminal end is a challenge due to the characteristics of e...

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Autores principales:	Qiang Zhang, Thomas J D Jørgensen, Peter E Nielsen, Niels Erik Møllegaard
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/54aa6160cde24412b38ae648c69d0f9a
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spelling	oai:doaj.org-article:54aa6160cde24412b38ae648c69d0f9a2021-11-18T08:29:30ZA phosphorylation tag for uranyl mediated protein purification and photo assisted tag removal.1932-620310.1371/journal.pone.0091138https://doaj.org/article/54aa6160cde24412b38ae648c69d0f9a2014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24599526/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Most protein purification procedures include an affinity tag fused to either the N or C-terminal end of the protein of interest as well as a procedure for tag removal. Tag removal is not straightforward and especially tag removal from the C-terminal end is a challenge due to the characteristics of enzymes available for this purpose. In the present study, we demonstrate the utility of the divalent uranyl ion in a new procedure for protein purification and tag removal. By employment of a GFP (green florescence protein) recombinant protein we show that uranyl binding to a phosphorylated C-terminal tag enables target protein purification from an E. coli extract by immobilized uranyl affinity chromatography. Subsequently, the tag can be efficiently removed by UV-irradiation assisted uranyl photocleavage. We therefore suggest that the divalent uranyl ion (UO22+) may provide a dual function in protein purification and subsequent C-terminal tag removal procedures.Qiang ZhangThomas J D JørgensenPeter E NielsenNiels Erik MøllegaardPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 3, p e91138 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Qiang Zhang Thomas J D Jørgensen Peter E Nielsen Niels Erik Møllegaard A phosphorylation tag for uranyl mediated protein purification and photo assisted tag removal.
description	Most protein purification procedures include an affinity tag fused to either the N or C-terminal end of the protein of interest as well as a procedure for tag removal. Tag removal is not straightforward and especially tag removal from the C-terminal end is a challenge due to the characteristics of enzymes available for this purpose. In the present study, we demonstrate the utility of the divalent uranyl ion in a new procedure for protein purification and tag removal. By employment of a GFP (green florescence protein) recombinant protein we show that uranyl binding to a phosphorylated C-terminal tag enables target protein purification from an E. coli extract by immobilized uranyl affinity chromatography. Subsequently, the tag can be efficiently removed by UV-irradiation assisted uranyl photocleavage. We therefore suggest that the divalent uranyl ion (UO22+) may provide a dual function in protein purification and subsequent C-terminal tag removal procedures.
format	article
author	Qiang Zhang Thomas J D Jørgensen Peter E Nielsen Niels Erik Møllegaard
author_facet	Qiang Zhang Thomas J D Jørgensen Peter E Nielsen Niels Erik Møllegaard
author_sort	Qiang Zhang
title	A phosphorylation tag for uranyl mediated protein purification and photo assisted tag removal.
title_short	A phosphorylation tag for uranyl mediated protein purification and photo assisted tag removal.
title_full	A phosphorylation tag for uranyl mediated protein purification and photo assisted tag removal.
title_fullStr	A phosphorylation tag for uranyl mediated protein purification and photo assisted tag removal.
title_full_unstemmed	A phosphorylation tag for uranyl mediated protein purification and photo assisted tag removal.
title_sort	phosphorylation tag for uranyl mediated protein purification and photo assisted tag removal.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/54aa6160cde24412b38ae648c69d0f9a
work_keys_str_mv	AT qiangzhang aphosphorylationtagforuranylmediatedproteinpurificationandphotoassistedtagremoval AT thomasjdjørgensen aphosphorylationtagforuranylmediatedproteinpurificationandphotoassistedtagremoval AT peterenielsen aphosphorylationtagforuranylmediatedproteinpurificationandphotoassistedtagremoval AT nielserikmøllegaard aphosphorylationtagforuranylmediatedproteinpurificationandphotoassistedtagremoval AT qiangzhang phosphorylationtagforuranylmediatedproteinpurificationandphotoassistedtagremoval AT thomasjdjørgensen phosphorylationtagforuranylmediatedproteinpurificationandphotoassistedtagremoval AT peterenielsen phosphorylationtagforuranylmediatedproteinpurificationandphotoassistedtagremoval AT nielserikmøllegaard phosphorylationtagforuranylmediatedproteinpurificationandphotoassistedtagremoval
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A phosphorylation tag for uranyl mediated protein purification and photo assisted tag removal.

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