Structure-Based Design and Synthesis of a New Phenylboronic-Modified Affinity Medium for Metalloprotease Purification
AbstractMetalloproteases are emerging as useful agents in the treatment of many diseases including arthritis, cancer, cardiovascular diseases, and fibrosis. Studies that could shed light on the metalloprotease pharmaceutical applications require the pure enzyme. Here, we reported the structure-based design and synthesis of the affinity medium for the efficient purification of metalloprotease using the 4-aminophenylboronic acid (4-APBA) as affinity ligand, which was coupled with Sepharose 6B via cyanuric chloride as spacer. The molecular docking analysis showed that the boron atom was interacting with the hydroxyl group of Ser176 residue, whereas the hydroxyl group of the boronic moiety is oriented toward Leu175 and His177 residues. In addition to the covalent bond between the boron atom and hydroxyl group of Ser176, the spacer between boronic acid derivatives and medium beads contributes to the formation of an enzyme-medium complex. With this synthesized medium, we developed and optimized a one-step purification procedure and applied it for the affinity purification of metalloproteases from three commercial enzyme products. The native metalloproteases were purified to high homogeneity with more than 95% purity. The novel purification method developed in this work provides new opportunities for scientific, industrial and pharmaceutical projects. View Full-Text
- Supplementary File 1:
Supplementary (DOCX, 513 KB)
Share & Cite This Article
Li, S.; Wang, L.; Xu, X.; Lin, S.; Wang, Y.; Hao, J.; Sun, M. Structure-Based Design and Synthesis of a New Phenylboronic-Modified Affinity Medium for Metalloprotease Purification. Mar. Drugs 2017, 15, 5.
Li S, Wang L, Xu X, Lin S, Wang Y, Hao J, Sun M. Structure-Based Design and Synthesis of a New Phenylboronic-Modified Affinity Medium for Metalloprotease Purification. Marine Drugs. 2017; 15(1):5.Chicago/Turabian Style
Li, Shangyong; Wang, Linna; Xu, Ximing; Lin, Shengxiang; Wang, Yuejun; Hao, Jianhua; Sun, Mi. 2017. "Structure-Based Design and Synthesis of a New Phenylboronic-Modified Affinity Medium for Metalloprotease Purification." Mar. Drugs 15, no. 1: 5.
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.