A comprehensive platform to investigate protein–metal ion interactions by affinity capillary electrophoresis

Abstract

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 6min for all of the investigated interactions. The method has been successfully applied for the interaction study of Li+, Na+, Mg2+, Ca2+, Ba2+, Al3+, Ga3+, La3+, Pd2+, Ir3+, Ru3+, Rh3+, Pt2+, Pt4+, Os3+, Au3+, Au+, Ag+, Cu1+, Cu2+, Fe2+, Fe3+, Co2+, Ni2+, Cr3+, V3+, MoO42− and SeO32− 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.