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 Citations (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.

Original language	English
Article number	e91138
Journal	PloS one
Volume	9
Issue number	3
Pages (from-to)	1-6
Number of pages	6
ISSN	1932-6203
DOIs	https://doi.org/10.1371/journal.pone.0091138
Publication status	Published - 5 Mar 2014

Access to Document

10.1371/journal.pone.0091138

Cite this

@article{ae4b1de39bb0431da0d9844c65f35e65,

title = "A Phosphorylation Tag for Uranyl Mediated Protein Purification and Photo Assisted Tag Removal",

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.",

author = "Qiang Zhang and J{\o}rgensen, {Thomas. J. D.} and Nielsen, {Peter E} and M{\o}llegaard, {Niels Erik}",

year = "2014",

month = mar,

day = "5",

doi = "10.1371/journal.pone.0091138",

language = "English",

volume = "9",

pages = "1--6",

journal = "PLoS Computational Biology",

issn = "1932-6203",

publisher = "Public Library of Science",

number = "3",

}

TY - JOUR

T1 - A Phosphorylation Tag for Uranyl Mediated Protein Purification and Photo Assisted Tag Removal

AU - Zhang, Qiang

AU - Jørgensen, Thomas. J. D.

AU - Nielsen, Peter E

AU - Møllegaard, Niels Erik

PY - 2014/3/5

Y1 - 2014/3/5

N2 - 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.

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.

U2 - 10.1371/journal.pone.0091138

DO - 10.1371/journal.pone.0091138

M3 - Journal article

C2 - 24599526

SN - 1932-6203

VL - 9

SP - 1

EP - 6

JO - PLoS Computational Biology

JF - PLoS Computational Biology

IS - 3

M1 - e91138

ER -

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

Abstract

Access to Document

Fingerprint

Cite this