Alhazmi Hassan A, Nachbar Markus, Albishri Hassan M, Abd El-Hady Deia, Redweik Sabine, El Deeb Sami, Wätzig Hermann
Institute of Medicinal and Pharmaceutical Chemistry, TU Braunschweig, D-38106 Braunschweig, Germany.
Chemistry Department, Faculty of Science, King Abdulaziz University, 80203 Jeddah, Saudi Arabia.
J Pharm Biomed Anal. 2015 Mar 25;107:311-7. doi: 10.1016/j.jpba.2015.01.017. Epub 2015 Jan 14.
In this work, the behavior of several metal ions with different globular proteins was investigated by affinity capillary electrophoresis. Screening was conducted by applying a proper rinsing protocol developed by our group. The use of 0.1M EDTA in the rinsing solution successfully desorbs metal ions from the capillary wall. The mobility ratio was used to evaluate the precision of the method. Excellent precision for repeated runs was achieved for different protein metal ion interactions (RSD% of 0.05-1.0%). Run times were less than 6 min for all of the investigated interactions. The method has been successfully applied for the interaction study of Li(+), Na(+), Mg(2+), Ca(2+), Ba(2+), Al(3+), Ga(3+), La(3+), Pd(2+), Ir(3+), Ru(3+), Rh(3+), Pt(2+), Pt(4+), Os(3+), Au(3+), Au(+), Ag(+), Cu(1+), Cu(2+), Fe(2+), Fe(3+), Co(2+), Ni(2+), Cr(3+), V(3+), MoO4(2-) and SeO3(2-) with bovine serum albumin, ovalbumin, β-lactoglobulin and myoglobin. Different interaction values were obtained for most of the tested metal ions even for that in the same metal group. Results were discussed and compared in view of metal and semimetal group's interaction behavior with the tested proteins. The calculated normalized difference of mobility ratios for each protein-metal ion interaction and its sign (positive and negative) has been successfully used to detect the interaction and estimate further coordination of the bound metal ion, respectively. The comprehensive platform summarizes all the obtained interaction results, and is valuable for any future protein-metal ion investigation.
在本研究中，通过亲和毛细管电泳研究了几种金属离子与不同球状蛋白的相互作用。采用本课题组开发的适当冲洗方案进行筛选。冲洗液中使用0.1M乙二胺四乙酸（EDTA）可成功从毛细管壁解吸金属离子。迁移率比用于评估该方法的精密度。对于不同的蛋白质-金属离子相互作用，重复运行具有出色的精密度（相对标准偏差为0.05-1.0%）。所有研究的相互作用的运行时间均小于6分钟。该方法已成功应用于锂离子（Li⁺）、钠离子（Na⁺）、镁离子（Mg²⁺）、钙离子（Ca²⁺）、钡离子（Ba²⁺）、铝离子（Al³⁺）、镓离子（Ga³⁺）、镧离子（La³⁺）、钯离子（Pd²⁺）、铱离子（Ir³⁺）、钌离子（Ru³⁺）、铑离子（Rh³⁺）、铂离子（Pt²⁺）、铂离子（Pt⁴⁺）、锇离子（Os³⁺）、金离子（Au³⁺）、亚金离子（Au⁺）、银离子（Ag⁺）、亚铜离子（Cu⁺）、铜离子（Cu²⁺）、亚铁离子（Fe²⁺）、铁离子（Fe³⁺）、钴离子（Co²⁺）、镍离子（Ni²⁺）、铬离子（Cr³⁺）、钒离子（V³⁺）、钼酸根离子（MoO₄²⁻）和亚硒酸根离子（SeO₃²⁻）与牛血清白蛋白、卵清蛋白、β-乳球蛋白和肌红蛋白的相互作用研究。即使对于同一金属组中的金属离子，大多数测试金属离子也获得了不同的相互作用值。根据金属和半金属组与测试蛋白质的相互作用行为对结果进行了讨论和比较。计算得到的每种蛋白质-金属离子相互作用的迁移率比归一化差值及其符号（正和负）已分别成功用于检测相互作用和估计结合金属离子的进一步配位情况。该综合平台总结了所有获得的相互作用结果，对未来任何蛋白质-金属离子研究都具有重要价值。
