Six To Be Adrenergic Drugs And Three Myosin Interaction And Its Applications,

Objective Three globulins (including human serum albumin,bovine serum albumin andlysozyme) were used as model proteins. Several adrenergic drugs were selected as researchtargets, such as ephedrine, pseudoephedrine, methylephedrine, terbutaline, salbutamol,clenbuterol. Under simulative physiological condition, the research and analytical application ofthe interaction between adrenergic drugs and three globulins were investigated withspectroscopy.Methods The changes of the UV-vis absorbance before and after the interaction of adrenergicdrugs with three globulins were studied by the ultraviolet difference spectra. The correlatingparameters and the interaction force of the interaction between adrenergic drugs with threeglobulins were obtained by the fluorescence spectroscopy and the influence of common metalions such as Cu²⁺, Mg²⁺, Co²⁺, Zn²⁺, Al³⁺, Ni²⁺ on the interaction of adrenergic drugs with threeglobulins was investigated.The effects of three globulins molecular conformation wereinvestigated by synchronous fluorescence. The content of human serum albumins in biologicalsample was determined by fluorescence spectrometry based on the interaction of adrenergicdrugs with human serum albumin, which provide a new method for the determination of humanserum albumin.Results The 1:1 compounds were formed between adrenergic drugs and metal ions. Theinteraction between adrenergic drugs and three globulins were studied by the ultravioletdifference spectra.The results of fluorescence spectrometry showed that the endogenous fluorescence of threeglobulins had been significantly quenched by adrenergic drugs.The mechanism of fluorescencequenching were static quenching with non-radiation energy transfer, the major driving forcewere hydrogen bond and Vander Waals. The binding sites of adrenergic drugs on three globulinswere one. The 1:1 complexes were formed between six adrenergic drugs and three globulins. Thebinding parameters of adrenergic drugs and BSA were as follows: the binding constants K were6.80×10³, 5.83×10³, 5.04×10³, 6.07×10³, 9.12×10³ and 7.06×10³ L·mol^-1, the binding distanceswere 0.323, 0.340, 0.363, 0.331, 0.215 and 0.241nm respectively. The binding parameters ofadrenergic drugs and HSA were as follows: the binding constants K were 4.38×10³, 4.23×10³,3.14×10³, 2.92×10³, 7.52×10³ and 5.87×10³L·mol^-1, the binding distances were 0.334, 0.350,0.369, 0.377, 0.241 and 0.250nm,respectively. The binding parameters of adrenergic drugs andLYSO were as follows: the binding constants K were 5.11×10³, 4.04×10³, 2.80×10³, 2.40×10³,7.89×10³ and 3.66×10³ L·mol^-1, the binding distances were 0.320, 0.322, 0.349, 0.396, 0.269 and0.332nm, respectively. The results of synchronous fluorescence demonstrated that the binding site was closer totryptophan residues. Three globulins molecular conformation was not changed by adrenergicdrugs.The content of human serum albumin in biological sample was determined by fluorescencebased on the interaction of adrenergic drugs with human serum albumin.The average recoveryrate of human serum albumin was 96.4±1.9%, 95.2±2.8%, 100±2.9%, 96.4±1.4%, 102±3.6%,96.1±2.5%, and the RSD was 2.0%, 3.0%, 2.9%, 1.5%, 3.5% and 2.6%, respectively.Conclusion Three globulins molecular conformation was basically unchanged, after sixadrenergic drugs interact three globulins. The binding constants were influenced by the metalions, but the major driving force had no change. The fluorescence method for determination ofhuman serum albumin in biological sample was simple and high sensitive.