Guowen Zhang

Interaction of alpinetin with bovine serum albumin: Probing of the mechanism and binding site by spectroscopic methods

The binding interaction between alpinetin and bovine serum albumin (BSA) in physiological buffer solution (pH 7.4) was investigated by fluorescence, UV-vis spectroscopy and Fourier transform infrared (FT-IR) spectroscopy. It was proved from fluorescence spectra that the fluorescence quenching of BSA by alpinetin was probably a result of the formation of BSA-alpinetin complexes, and the binding constant (K(a)) were determined according to the modified Stern-Volmer equation. The thermodynamic parameters, enthalpy change (DeltaH) and entropy change (DeltaS), were calculated to be 22.10kJmol(-1) and 166.04Jmol(-1)K(-1), respectively, which indicated that the interaction between alpinetin and BSA was driven mainly by hydrophobic interaction. Moreover, the competitive experiments of site markers suggested that the binding site of alpinetin to BSA was located in the region of subdomain IIA (sudlow site I). The binding distance (r) between the donor (BSA) and the acceptor (alpinetin) was 3.32nm based on the Förster theory of non-radioactive energy transfer. In addition, the results of synchronous fluorescence and FT-IR spectra demonstrated that the microenvironment and the secondary structure of BSA were changed in the presence of alpinetin.

Spectroscopic studies on the interaction between carbaryl and calf thymus DNA with the use of ethidium bromide as a fluorescence probe

The interaction between carbaryl and calf thymus DNA (ctDNA) was investigated under simulated physiological conditions (Tris-HCl buffer of pH 7.4) using ethidium bromide (EB) dye as a probe by UV-vis absorption, fluorescence and circular dichroism (CD) spectroscopy, as well as DNA melting studies and viscosity measurements. It can be concluded that carbaryl molecules could intercalate into the base pairs of DNA as evidenced by hyperchromic effect of absorption spectra, decreases in iodide fluorescence quenching effect, induced CD spectral changes, and significant increases in melting temperature and relative viscosity of DNA. The binding constants and thermodynamic parameters of carbaryl with DNA were obtained by the fluorescence quenching method. Furthermore, a chemometrics approach, parallel factor analysis (PARAFAC), was applied to resolve the measured three-way synchronous fluorescence spectral data matrix of the competitive interaction between carbaryl and EB with DNA, and the results provided simultaneously the concentration profiles and corresponding pure spectra for three reaction components (carbaryl, EB and DNA-EB complex) of the kinetic system at equilibrium. This PARAFAC analysis demonstrated the intercalation of carbaryl to the DNA helix by substituting for EB in the DNA-EB complex.

Spectroscopic studies of the interaction between pirimicarb and calf thymus DNA

The interaction between pirimicarb and calf thymus DNA in physiological buffer (pH 7.4) was investigated with the use of Neutral Red (NR) dye as a spectral probe by UV-vis absorption, fluorescence and circular dichroism (CD) spectroscopy, as well as viscosity measurements and DNA melting techniques. The results revealed that an intercalation binding should be the interaction mode of pirimicarb to DNA. CD spectra indicated that pirimicarb induced conformational changes of DNA. The binding constants of pirimicarb with DNA were obtained by the fluorescence quenching method. The thermodynamic parameters, enthalpy change (ΔHθ) and entropy change (ΔSθ) were calculated to be -52.13±2.04 kJ mol(-1) and -108.8±6.72 J mol(-1) K(-1) according to the vant Hoff equation, which suggested that hydrogen bonds and van der Waals forces might play a major role in the binding of pirimicarb to DNA. Further, the alternative least squares (ALS) method was applied to resolve a complex two-way array of the absorption spectra data, which provided simultaneously the concentration information for the three reaction components, pirimicarb, NR and DNA-NR. This ALS analysis indicated that the intercalation of pirimicarb into the DNA by substituting for NR in the DNA-NR complex.

Probing the binding mode of psoralen to calf thymus DNA.

The binding properties between psoralen (PSO) and calf thymus DNA (ctDNA) were predicted by molecular docking, and then determined with the use of UV-vis absorption, fluorescence, circular dichroism (CD) and Fourier transform infrared (FT-IR) spectroscopy, coupled with DNA melting and viscosity measurements. The data matrix obtained from UV-vis spectra was resolved by multivariate curve resolution-alternating least squares (MCR-ALS) approach. The pure spectra and the equilibrium concentration profiles for PSO, ctDNA and PSO-ctDNA complex extracted from the highly overlapping composite response were obtained simultaneously to evaluate the PSO-ctDNA interaction. The intercalation mode of PSO binding to ctDNA was supported by the results from the melting studies, viscosity measurements, iodide quenching and fluorescence polarization experiments, competitive binding investigations and CD analysis. The molecular docking prediction showed that the specific binding most likely occurred between PSO and adenine bases of ctDNA. FT-IR spectra studies further confirmed that PSO preferentially bound to adenine bases, and this binding decreased right-handed helicity of ctDNA and enhanced the degree of base stacking with the preservation of native B-conformation. The calculated thermodynamic parameters indicated that hydrogen bonds and van der Waals forces played a major role in the binding process.

Binding properties of herbicide chlorpropham to DNA: Spectroscopic, chemometrics and modeling investigations

The binding properties of chlorpropham (CIPC) to calf thymus DNA (ctDNA) were investigated in vitro by UV-vis absorption, fluorescence, circular dichroism (CD) and Fourier transform infrared (FT-IR) spectroscopy coupled with molecular modeling method. The results obtained from UV-vis absorption, fluorescence and CD spectroscopic methods as well as DNA viscosity and melting measurements indicated that the binding of CIPC to ctDNA was an intercalative mode. The FT-IR analysis and molecular modeling showed that CIPC mainly bound to guanine base of ctDNA. The association constant of the ctDNA-CIPC complex was determined to be in the order of 10(4) L mol(-1) by fluorescence titration. The calculated enthalpy change and entropy change suggested that hydrophobic forces and hydrogen bonds played prominent roles in the binding process. Furthermore, multivariate curve resolution-alternating least squares (MCR-ALS) approach was used to analyze the combined UV-vis absorption data matrix from the CIPC-ctDNA reaction system. The concentration profiles of the reaction components (CIPC, ctDNA and CIPC-ctDNA complex) and their pure spectra were successfully obtained to monitor the process of CIPC interaction with ctDNA. This study may contribute to the understanding of the CIPC-ctDNA interaction mechanism and toxicological effect of CIPC at the molecular level.

Mechanism and conformational studies of farrerol binding to bovine serum albumin by spectroscopic methods

The mechanism and conformational changes of farrerol binding to bovine serum albumin (BSA) were studied by spectroscopic methods including fluorescence quenching technique, UV-vis absorption, circular dichroism (CD) spectroscopy and Fourier transform infrared (FT-IR) spectroscopy under simulative physiological conditions. The results of fluorescence titration revealed that farrerol could strongly quench the intrinsic fluorescence of BSA through a static quenching procedure. The thermodynamic parameters enthalpy change and entropy change for the binding were calculated to be -29.92 kJ mol(-1) and 5.06 J mol(-1) K(-1) according to the vant Hoff equation, which suggested that the both hydrophobic interactions and hydrogen bonds play major role in the binding of farrerol to BSA. The binding distance r deduced from the efficiency of energy transfer was 3.11 nm for farrerol-BSA system. The displacement experiments of site markers and the results of fluorescence anisotropy showed that warfarin and farrerol shared a common binding site I corresponding to the subdomain IIA of BSA. Furthermore, the studies of synchronous fluorescence, CD and FT-IR spectroscopy showed that the binding of farrerol to BSA induced conformational changes in BSA.

Simultaneous spectrophotometric determination of atrazine and cyanazine by chemometric methods

A spectrophotometric method for the simultaneous determination of two herbicides, atrazine and cyanazine, is described for the first time based on their reaction with p-aminoacetophenone in the presence of pyridine in hydrochloric acid medium. The absorption spectra were measured in the wavelength range of 400-600 nm. The optimized method indicated that individual analytes followed Beers law in the concentration ranges for atrazine and cyanazine were 0.2-3.5 mg L(-1) and 0.3-5.0 mg L(-1), and the limits of detection for atrazine and cyanazine were 0.099 and 0.15 mg L(-1), respectively. The original and first-derivative absorption spectra of the binary mixtures were performed as a pre-treatment on the calibration matrices prior to the application of chemometric models such as classical least squares (CLS), principal component regression (PCR), partial least squares (PLS). The analytical results obtained by using these chemometric methods were evaluated on the basis of percent relative prediction error and recovery. It was found that the application of PCR and PLS models for first-derivative absorbance data gave the satisfactory results. The proposed methods were successfully applied for the simultaneous determination of the two herbicides in several food samples.

Binding properties of butylated hydroxytoluene with calf thymus DNA in vitro.

The binding properties of butylated hydroxytoluene (BHT) with calf thymus DNA (ctDNA) in simulated physiological buffer (pH 7.4) were investigated using ethidium bromide (EB) dye as a fluorescence probe by various spectroscopic techniques including UV-vis absorption, fluorescence, circular dichroism (CD), and Fourier transform infrared (FT-IR) spectroscopy along with ctDNA melting studies and viscosity measurements. It was found that the binding of BHT to ctDNA could decrease the absorption intensity of ctDNA, significantly increase melting temperature and relative viscosity of ctDNA, and induce the changes in CD spectra. Moreover, the competitive binding studies showed that BHT was able to displace EB from the bound ctDNA-EB complex. All the experimental results indicated that the binding mode between BHT and ctDNA was an intercalation. The association constants between BHT and ctDNA were evaluated to be (4.78±0.04)×10(3), (2.86±0.02)×10(3) and (1.80±0.04)×10(3) L mol(-)(1) at 298, 304, 310K, respectively. Further, the FT-IR analysis revealed that BHT was more prone to interact with adenine and thymine base pairs, and no significant conformational transition of ctDNA occurred. Thermodynamic analysis of the binding data showed that the binding process was primarily driven by hydrogen bonds and van der Waals forces, as the values of the enthalpy change and the entropy change were calculated to be -62.47±0.07kJ mol(-)(1) and -139.22±0.22J mol(-)(1) K(-)(1), respectively.