A Ni-NTA-based red fluorescence probe for protein labelling in live cells?
Journal of Materials Chemistry B Pub Date: 2017-01-09 DOI: 10.1039/C6TB02848A
Abstract
The great success of a His6-Ni-nitrilotriaceate (Ni-NTA) system in protein purification has inspired scientists to develop novel Ni-NTA based fluoresent probes for imaging of proteins in live cells. Despite a great deal of effort being made, only very few such probes can cross the cell membrane to label intracellular proteins. Herein, a new red fluorescence probe (Ni-NTA-AB) consisting of a Ni-NTA moiety, BODIPY and an arylazide was designed and synthesized to selectively label His6-tagged proteins. Guided by the Ni-NTA, the probe selectively binds to proteins with a genetically fused His6-tag exemplified by a DNA repair protein, xerodermapigmentosum group A (XPA122), leading to ca. 26-fold fluorescence enhancement. Ni-NTA-AB could enter into live bacterial cells to label intracellular His6-tagged proteins with the assistance of Tween 80 and also to quantify His6-tagged proteins in SDS-PAGE upon photoactivation of arylazide with a wide linear range (25–1000 ng). This study may provide a new horizon for the multiple-channel tracking and labelling of His6-tagged proteins.
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Journal Name：Journal of Materials Chemistry B
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CAS no.: 89640-58-4
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