Interaction Of Metal Ions And Biopolymers Studied With Microdialysis/HPLC

Microdialysis is a newly developed technology, which can be used for sampling and separating biological components. It is being widely used in the field of life science such as neurological, physiological, pharmacological and behavioral study.In this thesis, a method for studying the interaction of metal ions with biopolymer was proposed and developed by microdialysis sampling technique combined with high-performance liquid chromatography. This method was successfully applied to determine binding constants and binding sites of metal ions with human serum albumin, and it was proved to be a simple, fast and accurate method.The common binding sites and relative binding strength of metals to proteins could be confirmed. It was affirmed that there is a common secondary binding site on HSA and BSA for Cu (II), Zn (II), Ni (II) and Cd (II). The binding strength sequences of these four metal ions on the secondary binding site of HSA and BSA were determined, but the binding strength sequence of metal ions on HSA is different from that on BSA.This method was applied to study interaction of metal ion with metalloenzyme. The interaction of Superoxide Dismutase (Cu2Zn2SOD) with active center ions of Cu( II) and Zn( II) and Urease with active center ion of Ni (II), respectively, with added metal ions and their amino acid complexes were studied quantitatively.Obtained results showed that these added substances could induce or/and exchange with active center metal ions in metalloenzyme on some degrees. Some metal derivative metalloenzyme or/and apoenzyme appeared in the solution. Based on the relation between changing of the activity of metalloenzyme with contents of active center metal ion, a method was proposed for predicting if metal ion substituted metalloenzyme is active.