TY  - JOUR
AU  - Han, Jizhong 
AU  - Sun, Yunlong 
AU  - Chen, Yu 
AU  - Li, Haoran 
AU  - Liu, Mengmeng 
AU  - Zeng, Bin 
PY  - 2019
TI  - Influence of Tags on the Binding Affinity of Acyl-CoA Binding Protein
JF  - American Journal of Biochemistry and Biotechnology
VL  - 15
IS  - 1
DO  - 10.3844/ajbbsp.2019.13.22
UR  - https://thescipub.com/abstract/ajbbsp.2019.13.22
AB  - 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.