Recombinant expression and purification of human TATA binding protein using a chimeric fusion

The TATA binding protein (TBP) is the central core protein of the transcription factor II D that binds directly to the TATA box and therefore plays an integral part in eukaryotic transcription. This pivotal position of TBP is underlined by the vast number of interaction partners involved. Expression and purification of human TATA binding protein (hTBP) has remained a challenge due to protein instability and the protein loss during expression and purification involved. Here, we present a novel approach for high yield expression and purification of human TBP core (hTBPc) protein. Protein fold and activity are verified by nuclear magnetic resonance (NMR) spectroscopy and microscale thermophoresis (MST).

► A novel expression and purification approach for human TATA binding protein is proposed. ► Stable expression is facilitated by fusion with yeast TAND12. ► The chimeric fusion allows affinity purification due to yTAND12/hTBPc interaction.