@article {10.3844/ajbbsp.2019.13.22,
article_type = {journal},
title = {Influence of Tags on the Binding Affinity of Acyl-CoA Binding Protein},
author = {Han, Jizhong and Sun, Yunlong and Chen, Yu and Li, Haoran and Liu, Mengmeng and Zeng, Bin},
volume = {15},
number = {1},
year = {2019},
month = {Jan},
pages = {13-22},
doi = {10.3844/ajbbsp.2019.13.22},
url = {https://thescipub.com/abstract/ajbbsp.2019.13.22},
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.},
journal = {American Journal of Biochemistry and Biotechnology},
publisher = {Science Publications}
}