Spectroscopy Studies On The Interaction Between Serum Protein And Small Molecules Drugs With Dye As Probe

This paper mainly studies 3 kinds of dyes and BSA interactions, and used these dyes as probes to study the specific binding reaction of several drugs with BSA respectively. The research content has been divided into five chapters:Chapter one: The research content, the research method on the interaction between small drug molecules and protein molecules were reviewed, and the types and the development prospect of fluorescent probe were introduced. A total of 106 references are cited.Chapter two: This article studied the combination of KM(or SM) and BSA with Eosin Y as a fluorescence probe, using the method of fluorescence spectrum. Results show that in the Tris-HCl buffer solution with the p H=7.40, EY and BSA made BSA occur fluorescence quenching, and its quenching mechanism was static quenching, and under different temperatures the binding constant, binding points, combined with distance and reaction type between EY with BSA were obtained. In this sense, it can be concluded that KM(or SM) specifically binds to Bovine serum albumin, and it determines the binding site on sub-domain IIA(site I) of KM(or SM) and BSA.Chapter three: Using Acriflavine(or Methyl Green) as fluorescent probe, seven kinds of drugs responding to the combination of the BSA were studied. The drugs are Streptomycin sulphate(SM), Kanamycin sulfate(KM), Gentamicin(GM), Amikacin(AM), Neomycin(NM), Thiamphenicol(TN), and Florfenicol(FN). And after reaction of BSA and these drugs, the fluorescence intensity of BSA has no obvious change. The research has shown that in Tris-HCl buffer solution with p H=7.40, ACF(or MG) made BSA fluorescence quenching, and its quenching mechanism belonged to static quenching, and the binding constant, binding points and reaction type of ACF(or MG) between BSA were obtained under different temperature and so on. Seven kinds of drugs after joined the BSA-ACF(or MG) system, respectively, and made the fluorescence intensity of BSA recovering. The degree of recovery were GM> TN> NM> FN> KM> SM> AM(or NM> AM> GM> SM> FN> KM> TN). This suggest that ACF(or MG), drugs, and the competition between BSA binding reaction, its reaction mechanism was discussed. Confirmed that with ACF(or MG) as A fluorescence probe, fluorescence spectrometry was used to study, you can know the 7 kinds of specific binding reaction happened between drugs and BSA, and used with the markers of specific binding to determine the 7 kinds of drugs on BSA bonding sites for IIA of the siteⅠ.Chapter four: In the Tris-HCl buffer solution with the p H=7.40, reaction mechanism between ACF, MG and BSA was studied at 293 K, 303 K and 310 K temperature by utilizing fluorescence quenching method and synchronous fluorescence method, respectively. The results indicated that ACF(or MG) could quench the intrinsic fluorescence of BSA strongly, and the quenching mechanism was a static quenching process; the electrostatic force played an important role on the conjugation reaction between ACF and BSA; the hydrogen bonding and van der Waals force played an important role on the conjugation reaction between MG and BSA. The order of magnitude of binding constants(Ka) was 104, the number of binding site(n) in the binary system was approximately equal to 1 and the primary binding for Hydrogen bonding and van der Waals force were located at the structure domain II A of BSA. The results obtained by the two methods were consistent, which indicated synchronous fluorescence spectroscopy can replace traditional fluorescence quenching method to study reaction mechanism of dyes with proteins.Chapter five: In Tris-HCl buffer solution(p H=7.40), the Moxifloxacin, Norfloxacin, Ofloxacin, Enrofloxacin, Lomefloxacin, Ciprofloxacin interaction with BSA were studied, respectively. Using the Scatchard equation correction equation, Lineweaver-Burk double reciprocal equation, double logarithmic equation, modified Stern-Volmer equation, and improved double logarithmic equation to calculate the binding constants and binding points of drugs and bovine serum albumin, respectively. Comparing the calculated results, indicated that the drug-protein binding reaction was 1:1, the calculated results of other equations very close except modified Scatchard equation results, where the improved double logarithmic equation calculate the number of results more reasonable and accurate.