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

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 one

JF - PloS one

IS - 3

M1 - e91138

ER -