A Phosphorylation Tag for Uranyl Mediated Protein Purification and Photo Assisted Tag Removal

Qiang Zhang; Thomas. J. D. Jørgensen; Peter E Nielsen; Niels Erik Møllegaard

doi:10.1371/journal.pone.0091138

A Phosphorylation Tag for Uranyl Mediated Protein Purification and Photo Assisted Tag Removal

Qiang Zhang, Thomas. J. D. Jørgensen, Peter E Nielsen, Niels Erik Møllegaard

Transcription, RNA, and Gene Medicine Program

5 Citationer (Scopus)

Abstract

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.

Originalsprog	Engelsk
Artikelnummer	e91138
Tidsskrift	PloS one
Vol/bind	9
Udgave nummer	3
Sider (fra-til)	1-6
Antal sider	6
ISSN	1932-6203
DOI	https://doi.org/10.1371/journal.pone.0091138
Status	Udgivet - 5 mar. 2014

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10.1371/journal.pone.0091138

Citationsformater

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abstract = "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.",

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AU - Zhang, Qiang

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AU - Nielsen, Peter E

AU - Møllegaard, Niels Erik

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AB - 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.

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A Phosphorylation Tag for Uranyl Mediated Protein Purification and Photo Assisted Tag Removal

Abstract

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