Study On The Interaction Of Proteins With Small Molecules And Determination Of Protein By Molecular Spectroscopies

We all know that protein is one of the basic materials to constitute life. As a main supporter of varieties activities of life, protein plays a very important role in the life activities and almost all of the processes in lives and takes on amounts of physiological functions, including immune and enzymatic and so on. Therefore, further research about protein has a positive significance to uncover the secrets of lives. And it will be one of the hottest research points in the fields of clinical physics, chemistry and life sciences during the 21st century.As two varieties of important proteins, both trypsin and hemoglobin are really ire-placeable in the activities of life.In this thesis, a new method to determine hemoglobin has been established and the interaction mechanisms of drugs with hemoglobin and trypsin have been investigated by spectroscopic technologies and laws related. We have done these all above to provide some important information about structure and functions of protein, meanwhile some reliable data in pathology and medicine designing has been gained, too.The main content of the thesis is as follows:1. The significance and development in the research of protein have been outlined.2. Under the acid condition, hemoglobin has been oxidated to be a substance of strong fluorescence by the cerium-ion (Ⅳ). Based on that, a new method of determination of hemoglobin has been created and used to determine hemoglobin in human serum. The calibration graph is linear in the range of 1.0×10-7～8.0×10-7 mol·L-1 with the detection limit of 1.0×10-9 mol·L-1,and recovering rate is 96.80%～102.6%.3. Under physiological condition, we have investigated the interaction mechanisms of antibiotic drugs with bovine hemoglobin by means of fluorescence spectroscopy and UV-vis absorption spectroscopy. The associating constants have been calculated and the properties of interaction have been studied by the thermodynamic parameters. Meanwhile, the distance r between the acceptor and the donor has been determined according to Forster non-radiation energy transfer theory. The effect of antibiotic drugs on the conformation of hemoglobin has been analyzed by synchronous fluorescence spectra.4. That the spectrum of trypsin interacted with caffeine and theophylline was changed has been researched. The results have been got. First, the quenching of alkaloid with trypsin were both dynamic; Second, energy transfer should be existed via Forster non-radiation energy transfer theory synchronous fluorescence spectra; Third, the conformation was changed.5.The spectrum properties of trypsin have been varied after the interaction of trypsin and SCX[4,6,8]. Based on that, some conclusions have been exactly got. The quenching was dynamic, and the acting forces were all hydrophobic interaction. Expectedly, the configuration was changed as well.