Bioengineering Department, Hacettepe University, Ankara, Turkey.
Department of Environmental Engineering, Hacettepe University, Ankara, Turkey.
J Mol Recognit. 2021 Feb;34(2):e2875. doi: 10.1002/jmr.2875. Epub 2020 Sep 4.
Immobilized metal ion affinity chromatography (IMAC) has become a widespread analytical and preparative separation method for therapeutic proteins, peptides nucleic acids, hormones, and enzymes. N-Methacryloyl-l-histidine Methyl Ester (MAH) monomer is recently used as a synthesized affinity ligand in IMAC. It is capable of chelating with many transition metal ions such as Zn , Ni , and Cu ions through its histidine residue. In this way, proteins can bind selectively to these immobilized metal ions through MAH as a ligand in affinity chromatography. In this study, we applied the computational docking method on the interactions that occur between the MAH monomer and its complexes with Zn ions as ligands and protein molecules as targets. MAH monomer was drawn and created using the Avogadro software as an optimization tool. Human insulin (Ins) molecule and horse heart cytochrome C (Cyt C) were selected as target proteins to interact with MAH monomer as affinity ligand. Automated docking software AutoDock v4.2 was used for docking of MAH monomer to Ins and Cyt C, respectively. The affinity ligand complexes with Zn ions bound to one, two, and three moles of MAH were studied and compared separately. The lowest binding energies of Ins and Cyt C proteins in 1:1 mol ratio of MAH-Zn were found as (-4.14) and (-4.92) kcal/mol, respectively.
固定化金属离子亲和层析（IMAC）已成为治疗性蛋白质、肽、核酸、激素和酶的广泛分析和制备分离方法。N-甲基丙烯酰-L-组氨酸甲酯（MAH）单体最近被用作 IMAC 中的合成亲和配体。它能够通过其组氨酸残基与许多过渡金属离子（如 Zn、Ni 和 Cu 离子）螯合。通过这种方式，蛋白质可以通过 MAH 作为亲和层析中的配体选择性地结合到这些固定化金属离子上。在这项研究中，我们应用计算对接方法研究 MAH 单体与其与 Zn 离子作为配体的复合物以及蛋白质分子作为靶标的相互作用。MAH 单体使用 Avogadro 软件作为优化工具进行绘制和创建。选择人胰岛素（Ins）分子和马心细胞色素 C（Cyt C）作为靶蛋白，与 MAH 单体作为亲和配体相互作用。使用 AutoDock v4.2 自动对接软件分别对接 MAH 单体与 Ins 和 Cyt C。分别研究并比较了与 Zn 离子结合的 MAH 单体结合一个、两个和三个摩尔的复合物。在 MAH-Zn 的 1:1 mol 比下，Ins 和 Cyt C 蛋白质的最低结合能分别为(-4.14)和(-4.92) kcal/mol。
