Research Article Open Access

Influence of Tags on the Binding Affinity of Acyl-CoA Binding Protein

Jizhong Han1, Yunlong Sun1, Yu Chen1, Haoran Li1, Mengmeng Liu1 and Bin Zeng1
  • 1 Jiangxi Science and Technology Normal University, China

Abstract

Tagged fusion proteins are frequently employed for protein purification methods, but their effects on protein function and binding affinity are rarely studied. Here we expressed recombinant protein Acyl-CoA Binding Protein (ACBP) cloned from the full-length cDNA of Aspergillus oryzae and Saccharomyces cerevisiae. ACBP was expressed in Escherichia coli fused to a Maltose-Binding Protein (MBP) and Histidine-tag fusion. Recombinant ACBP was purified using affinity chromatography columns and high protein purity was achieved. Microscale Thermophoresis (MST) binding assays showed that recombinant AoAcbp1 had a greater affinity for Palmitoyl-CoA (Kd = 35 nM) and Stearoyl-CoA (Kd = 23 nM) whilst recombinant ScAcbp had a greater affinity for Myristoyl-CoA (Kd = 31 nM) and Palmitoyl-CoA (Kd = 51 nM). In addition, MBP tagged ACBP had comparable binding affinities to His-tagged ACBP. Taken together, these data highlight that the size of the tagged fusion protein does not influence protein ACBP binding affinity.

American Journal of Biochemistry and Biotechnology
Volume 15 No. 1, 2019, 13-22

DOI: https://doi.org/10.3844/ajbbsp.2019.13.22

Submitted On: 1 August 2018 Published On: 23 January 2019

How to Cite: Han, J., Sun, Y., Chen, Y., Li, H., Liu, M. & Zeng, B. (2019). Influence of Tags on the Binding Affinity of Acyl-CoA Binding Protein. American Journal of Biochemistry and Biotechnology, 15(1), 13-22. https://doi.org/10.3844/ajbbsp.2019.13.22

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Keywords

  • Acyl-CoA Binding Protein
  • Tagged Fusion Protein
  • Affinity Chromatography
  • Microscale Thermophoresis
  • Affinity Analysis