Study On Interaction Of Proteins With Nucleic Acids And Small Molecules And Its Analytical Application By Resonance Rayleigh Scattering Spectra

In recent ten years, the interaction between protein and nucleic acid has attracted more and more attention because of its important role in physiology. Many analytical methods have been used in the mechanical studies of the interaction and recognition between the two biological macromolecules. For its high sensitivity to effect of noncovalence, Resonance Rayleigh scattering (RRS), such as association, aggregation and the dipole-dipole of molecules, can be used in the study of mechanism and detection. In the article, the characteristic, the mechanism of the reaction between protein and nucleic acid by RRS method, and the method can also be used to detect protein and nucleic acid.Besides, the spectral characteristics, the affecting factors and the properties of analytical chemistry, the reaction mechanism and their analytical applications have been studied. In addition, we also determined protein with tannic acid and DNA with some metal-ion chelate.1 Absorption spectra study on the interaction of protein with some DNAThe absorption spectra of interaction between protein and nucleic acid were studied, and it can be used to research the mechanism of reaction. The results showed that the absorption spectra changed as soon as the two biomacromolecules reacted with each other, and it can give some information of the interaction. Base on the reaction, a novel method of protein and nucleic acid can be set up. When DNA were determined, the detect limitswere between 191.4 ng and 276.4 ng, ·mL^-1 but the sensitivity to RNA was lower. When we determined the proteins by hsDNA, the detect limits were 27.4 μg ·mL^-1HSA), 2.0 μg ·mL^-1 (Papain), 4.1 μg ·mL^-1Pepsin), 2.0μg ·mL^-1(BSA) and 28 μg ·mL^-1 (Trypsin), respectively.2 RRS study on interaction of trypsin with some DNAThe interaction between Trypsin and DNA was studied. The optimal interaction conditions, the affecting factors, the properties of analytical chemistry of the binding complex were investigated and the binding ratio of trypsin with DNA. The binding mode, binding ratio of trypsin with DNA and the reasons of RRS enhanced has been a preliminary discussion. The interaction can also be used to detect protein and nucleic acid. When DNA was detect by trypsin, the detect limits were between 0.42¹.06 ng-mL"1.3 RRS study of interaction of protein with some DNAThe RRS spectra of hsDNA reacted with Human serum albumin, bovine serum albumin, papain, pepsin, hemoglobin and y-globin were studied. The results showed that the reaction of hsDNA with proteins can cause the enhancement of Ims- The characteristic of RRS spectra and the reaction condition were experimented. Associating with other molecular spectra, the binding mode was also discussed. When we determined proteins with hsDNA, the linear range were 0.05^<sup>12 ^g*mL-1 (HSA), 0.04 15 /jgvnL'^BSA) , 0.1 20/wg?mL'1 (y-globin) and 0.1 lfyfgTnL"1 (hemoglobin), and the detect limit were from 12.6 ng'mL"1 to 43.4 ng^mL"1 o When we determined hsDNA with proteins, the linear range were 0.4x10 "3¹.5 ftg'mL'1 (HSA), O.3xlO"3¹.6 ^g-mL"1 (BSA), 0.1xl0'3².7 //g'mL'1 (Papain), O.4xlO"3².5 /ig'rnL"1 (Pepsin), O.2xlO"3².8 /ig'mL"1 (y-globin) and 1.4xlO'3¹.9 (hemoglobin), the detect limits were 1.2 ng-mL"1 (HSA), 0.7 ng'mL"1 (BSA), 0.3 ng'mL'1 (Papain) 1.1 ng'mL"1 (Pepsin) O^ng^mL'1 (yglobin) and 0.4 ng^mL"1 (hemoglobin).4 RRS method for the determination of protein with tannic acidWe studied the spectra characteristics of tannic acid with HSA and BSA and some other proteins, and found out that the intensity of RRS extensively enhanced by the reaction between the albumin and tannic acid. The spectra characteristics, the effect factors, the optimum conditions and the binding mode of the reaction had been investigated. The method can be used to detect BSA and HSA. When we determined BSA, the intensity of RRS spectra directly proportional to the concentration of protein between 0.04¹00 /Ug'mL"1, The detect limit was 12.8 ngTnL"1. The linear range of HSA was 0.03⁹0 ^gvnL"1, and the detect limit was 10.4 ng#mL"\ Under the experiment condition, the signal of the other protein react with tannic acid were lower, so the method can be used to detect albumin with the presence of other proteins. We detect the amount of blood serum samples, and the result were accord with standard method.5 RRS method for the determination of DNA with metal ion-BDHAchelatesThe article studied the RRS spectra characteristics of the ternary complexes of BDNA associated with Cu2+, Al3+, Cd2+ and DNA. The results showed that theconformation of the ternary complex would cause the enhancement of Iurs- The three complexes had similar spectra. The maximum peaks were at 357nm the spectra characteristics, and the two lower peaks were at 310 nm and 537nm. The effect factors, the optimum conditions of the reaction, the influence of foreign substances and the reaction mechanism have been investigated. When we detect hsDNA with BDHA-metal ion complex, the detect limits are 2.0 ng'mL"1 (BDHA-Cu2 + ), SS.Sng'mL"1 (BDHA-A13+) and TT.Sng'mU^BDHA-Cd2*), respectively.