Biomedical subjects

J F Neault

Publications and source records attributed to J F Neault.

11 recordsLinked to original sources

PubMed2000

Interaction of taxol with human serum albumin.

Taxol (paclitaxel) is an anticancer drug, which interacts with microtuble proteins, in a manner that catalyzes their formation from tubulin and stabilizes the resulting structures (Nogales et al., Nature 375 (1995) 424-427). This study was designed to examine the interaction of taxol with human serum albumin (HSA) in aqueous solution at physiological pH with drug concentrations of 0.0001-0.1 mM, and HSA (fatty acid free) concentration of 2% w/v. Gel electrophoresis, absorption spectra and Fourier transform infrared (FTIR) spectroscopy with self-deconvolution and second-derivative resolution enhancement were used to determine the drug binding mode, binding constant and the protein secondary structure in the presence of taxol in aqueous solution. Spectroscopic evidence showed that taxol-protein interaction results into two types of drug-HSA complexes with overall binding constant of K=1.43 x 10(4) M(-1). The molar ratios of complexes were of taxol/HSA 30/1 (30 mM taxol) and 90/1 (90 mM taxol) with the complex ratios of 1.9 and 3.4 drug molecules per HSA molecule, respectively. The taxol binding results in major protein secondary structural changes from that of the alpha-helix 55 to 45% and beta-sheet 22 to 26%, beta-anti 12 to 15% and turn 11 to 16%, in the taxol-HSA complexes. The observed spectral changes indicate a partial unfolding of the protein structure, in the presence of taxol in aqueous solution.

Drug Interactions↗

PubMed1999

Structural analysis of DNA-chlorophyll complexes by Fourier transform infrared difference spectroscopy.

Porphyrins and metalloporphyrins are strong DNA binders. Some of these compounds have been used for radiation sensitization therapy of cancer and are targeted to interact with cellular DNA. This study was designed to examine the interaction of calf thymus DNA with chlorophyll a (CHL) in aqueous solution at physiological pH with CHL/DNA(phosphate) ratios (r) of 1/160, 1/80, 1/40, 1/20, 1/10, and 1/5. Fourier transform infrared (FTIR) difference spectroscopy was used to characterize the nature of DNA-pigment interactions and to establish correlations between spectral changes and the CHL binding mode, binding constant, sequence selectivity, DNA secondary structure, and structural variations of DNA-CHL complexes in aqueous solution. Spectroscopic results showed that CHL is an external DNA binder with no affinity for DNA intercalation. At low pigment concentration (r = 1/160, 1/80, and 1/40), there are two major binding sites for CHL on DNA duplex: 1) Mg-PO2 and 2) Mg-N7 (guanine) with an overall binding constant of K = 1.13 x 10(4) M-1. The pigment distributions are 60% with the backbone PO2 group and 20% with the G-C base pairs. The chlorophyll interaction is associated with a major reduction of B-DNA structure in favor of A-DNA. At high chlorophyll content (r = 1/10), helix opening occurs, with major spectral alterations of the G-C and A-T bases. At high chlorophyll concentration (1/5), pigment aggregation is observed, which does not favor CHL-DNA complexation.

Animals↗

PubMed1999

Interaction of cisplatin drug with RNase A.

cis-Pt(NH3)2Cl2 (cisplatin) is an antitumor drug with many severe toxic side effects including enzymatic structural changes associated with its mechanism of action. This study is designed to examine the interaction of cisplatin drug with ribonuclease A (RNase A) in aqueous solution at physiological pH, using drug concentration of 0.0001 mM to 0.1 mM with final protein concentration of 2% w/v. Absorption spectra and Fourier transform infrared (FTIR) spectroscopy with its self-deconvolution, second derivative resolution enhancement and curve-fitting procedures were used to characterize the drug binding mode, association constant and the protein secondary structure in the cisplatin-RNase complexes. Spectroscopic results show that at low drug concentration (0.0001 mM), no interaction occurs between cisplatin and RNase, while at higher drug concentrations, cisplatin binds indirectly to the polypeptide C=O, C-N (via H2O or NH3 group) and directly to the S-H donor atom with overall binding constant 5.66 x 10(3)M(-1). At high drug concentration, major protein secondary structural changes occur from that of the alpha-helix 29% (free enzyme) to 20% and beta-sheet 39% (free enzyme) to 45% in the cisplatin-RNase complexes. The observed structural changes indicate a partial protein unfolding in the presence of cisplatin at high drug concentration.

Animals↗

PubMed1998

Interaction of cisplatin with human serum albumin. Drug binding mode and protein secondary structure.

Cis-diamminedichloroplatinum(II) (cisplatin) is an antitumor drug, which forms intrastrand cross-links DNA adducts. Protein interaction with cisplatin-DNA complexes induces DNA bending and biopolymer structural changes. This study is designed to examined the interaction of cisplatin with human serum albumin (HSA) in aqueous solution at physiological pH with drug concentrations of 0.0001 mM to 0.1 mM, and HSA (fatty acid free) concentration of 2% w/v. Absorption spectra and Fourier transform infrared (FTIR) spectroscopy with its self-deconvolution and second derivative resolution enhancement, as well as curve-fitting procedures, were used to determine the drug binding mode, drug binding constant and the protein secondary structure in aqueous solution. Spectroscopic evidence showed that at low drug concentration (0.0001 mM), minor cisplatin-protein interaction occurs, while at higher drug content (0.001 mM), major Pt-HSA complexation takes place via protein C=O, C-N and S-H donor groups with overall binding constant K = 8.52 x 10(2) M-1. At high drug concentration, cisplatin binding results in major protein secondary structural changes from that of the alpha-helix 55% (free HSA) to 45% and beta-sheet 22% (free HSA) to 32%, in the cisplatin-HSA complexes. The observed spectral changes indicate a partial unfolding of the protein structure, in the presence of cisplatin at high drug concentrations.

Binding Sites↗

PubMed1998

RNA-ascorbate interaction.

Ascorbic acid and divalent iron salts have been widely used to investigate the effects of reactive oxygen species in different biological targets such as nucleic acids, proteins and lipids. This study was designed to examine the interaction of yeast RNA with vitamin C in aqueous solution at physiological pH with drug/RNA(P)(P=phosphate) molar ratios of r=1/80, 1/40, 1/20, 1/10, 1/4 and 1/2. Absorption spectra and Fourier transform infrared (FTIR) difference spectroscopy were used to determine the ascorbate binding mode, binding constant, sequence selectivity and RNA secondary structure in aqueous solution. Spectroscopic evidence showed that at low drug concentration (r=1/80 and 1/40), no major ascorbate-RNA interaction occurs, while at higher drug concentrations (r>1/40), a major drug-RNA complexation was observed through both G-C and A-U base pairs and the backbone phosphate groups with k=31.80 M(-1). Evidence for this comes from large perturbations of the G-C vibrations at 1698 and 1488 cm(-1) and the A-U bands at 1654 and 1608 cm(-1) as well as the phosphate antisymmetric stretch at 1244 cm(-1). At r>1/10, minor structural changes occur for the ribose-phosphate backbone geometry with RNA remaining in the A-family structure. The drug distributions around double helix were about 55% with G-C, 33% A-U and 12% with PO2 groups. A comparison between ascorbate-RNA and ascorbate-DNA complexes showed minor differences. The ascorbate binding (H-bonding) is via anion CO and OH groups.

Ascorbic Acid↗

PubMed1997

RNA-diethylstilbestrol interaction studied by Fourier transform infrared difference spectroscopy.

Diethylstilbestrol (DES), a synthetic estrogen, is known to be a carcinogen in human and in animals. This study was designed to examine the interaction of DES with yeast RNA in aqueous solution at physiological pH with drug/RNA-phosphate (P) molar ratios of 1/80, 1/40, 1/20, 1/10, 1/4, and 1/2. Fourier transform infrared (FTIR) difference spectroscopy was used to determine the drug binding mode, the binding constant, the sequence selectivity, and RNA secondary structure in the RNA.DES complexes. Spectroscopic evidence showed that at low drug concentration (1/80 and 1/40), DES is intercalating through both Gua-Cyt and Ade-Urd base pairs with minor interaction with the backbone PO2 group (external binding). The calculated binding constant of K approximately 8.5 x 10(4) M-1 at a drug concentration of 3.12 x 10(-4) M shows that DES is a weaker intercalator than those of the methylene blue, acridine orange, and ethidium bromide. At high drug content (r > 1/40, where r represents the DES/RNA-phosphate molar ratio), a partial helix destabilization occurs with no alteration of RNA conformation upon drug complexation. However, a comparison with DNA.DES complexes showed that drug intercalation causes major reduction of the B-DNA structure in favor of A-DNA with no participation of the backbone PO2 group in the DES. DNA complexation.

DNA↗

PubMed1996

Diethylstilbestrol-DNA interaction studied by Fourier transform infrared and Raman spectroscopy.

The interaction of diethylstilbestrol (DES) with calf thymus DNA was investigated at physiological pH with drug/DNA (phosphate) molar ratios (r) of 1:40, 1:20, 1:10, 1:4, 1:2, and 1. Fourier transform infrared and laser Raman difference spectroscopy were used to establish correlations between spectral changes and drug binding mode, sequence selectivity, DNA conformation, and structural properties of DES.DNA complexes in aqueous solution. Spectroscopic results indicated that DES is a weak intercalator with affinity for A-T-rich regions. It is also a groove binder with a major interaction with the thymine O-2 atom. At low drug concentration (r = 1:40), the A-T-rich region is the main target of drug intercalation, while at a higher drug content (r > 1:5), external binding to the G-C bases also occurs with a partial helix destabilization. Evidence for this comes from the spectral alterations of the A-T vibrational frequencies at 1661 cm-1 (Raman) and 1663 and 1609 cm-1 (IR) and of the G-C vibrations at 1581 and 1491 cm-1 (Raman) and 1717 and 1492 cm-1 (IR). Drug intercalation leads to a major reduction of B-DNA structure in favor of A-DNA.

DNA↗

PubMed1996

Aspirin-DNA interaction studied by FTIR and laser Raman difference spectroscopy.

The interaction of calf-thymus DNA with aspirin is investigated in aqueous solution at pH 7-6 with drug/DNA (phosphate) molar ratios of r = 1/40, 1/20, 1/10, 1/5, 1/2, 1 and 2. Fourier transform infrared (FTIR) and laser Raman difference spectroscopy are used to determine drug binding sites, sequence preference and DNA secondary structure, as well as the structural variations of aspirin-DNA complexes in aqueous solution. Spectroscopic evidence showed that at low aspirin concentration (r =1/40), drug-DNA interaction is mainly through the backbone PO2 groups and the A-T base pairs. Such interaction largely perturbs the phosphate vibration at 1222 cm(-1) and the A-T bands at 1663 and 1609 cm(-1) with no major helix destabilization. At higher drug concentration (r > 1/20), the participation of the G-C bases in drug-DNA complexation was evident by strong perturbations of the guanine and cytosine vibrations at 1717 and 1494 cm(-1), with a partial helix destabilization. A major alteration of the B-DNA structure towards A-DNA occurs on drug complexation. The aspirin interaction was through anion CO and COOCH3 donor atoms with those of the backbone PO2 group and DNA bases donor sites (directly or indirectly via H2O molecules).

Animals↗

PubMed1996

An FTIR spectroscopic study of calf-thymus DNA complexation with Al(III) and Ga(III) cations.

The interaction of calf-thymus DNA with trivalent Al and Ga cations, in aqueous solution at pH = 6-7 with cation/DNA(P) (P = phosphate) molar ratios (r) 1/80, 1/40, 1/20, 1/10, 1/4 and 1/2 was characterized by Fourier Transform infrared (FTIR) difference spectroscopy. Spectroscopic results show the formation of several types of cation-DNA complexes. At low metal ion concentration (r = 1/80, 1/40), both cations bind mainly to the backbone PO2 group and the guanine N-7 site of the G-C base pairs (chelation). Evidence for cation chelate formation comes from major shifting and intensity increase of the phosphate antisymmetric stretch at 1222 cm-1 and the mainly guanine band at 1717 cm-1. The perturbations of A-T base pairs occur at high cation concentration with major helix destabilization. Evidence for cation binding to A-T bases comes from major spectral changes of the bands at 1663 and 1609 cm-1 related mainly to the thymine and adenine in-plane vibrations. A major reduction of the B-DNA structure occurs in favor of A-DNA upon trivalent cation coordination.

Aluminum↗

PubMed1995

Does DNA acid fixation produce left-handed Z structure?

The effects of acetic acid (HCOOCH3) on the solution structure of calf-thymus DNA are studied at pH 7.3-2.5 with acid/DNA(P) (phosphate) molar ratios (r) of 1/40, 1/20, 1/10, 1, 2, 10, 20 and 40. Fourier Transform infrared (FTIR) difference spectroscopy is used to establish correlations between spectral changes and base protonation, DNA conformational transition and structural variations of the acid-DNA complexes in aqueous solution. The FTIR difference spectroscopic results showed that protonation of cytosine and subsequent unpairing of the G-C base pairs begins at pH 4-3 and continues up to pH 2.5, where a complete base separation and base unstacking occur. Similarly, protonation of A-T base pairs starts at pH 4-3 and is completed at pH 2.5, where base separation and base unstacking are observed. The protonation of the G-C base pair leads to the formation of Hoogsteen-type H-bonding, before a complete G-C disruption. The biopolymer protonation leads to the formation of several non-B-DNA structures, including left-handed Z conformation.

Animals↗

PubMed1995

DNA-drug interaction. The effects of vitamin C on the solution structure of Calf-thymus DNA studied by FTIR and laser Raman difference spectroscopy.

The interaction of calf-thymus DNA with L-ascorbic acid was investigated in aqueous solution at pH=7.6 with drug/DNA(P)(P=phosphate) molar ratios (r) of 1/40, 1/20, 1/10, 1/5, 1/2, 1 and 2. Fourier Transform infrared (FTIR) and laser Raman difference spectroscopic techniques were used to establish correlations between spectral modifications and drug binding mode, sequence specificity, DNA melting and conformational changes, as well as structural variations of drug-DNA complexes in aqueous solution. Infrared and Raman spectroscopic results showed that at low drug concentration (r = 1/40), a B to A-type conformational conversion occurs with minor drug-DNA interaction through A-T bases. At r=1/20, drug-PO2 binding was also observed with reduced intensity of DNA inplane vibrational frequencies, due to the increased base-stacking interaction and duplex stability. At r> 1/20, major perturbations of DNA bases were observed for both A-T and G-C base pairs in the major and minor grooves of the duplex. Evidence for this comes form the shift of the infrared and Raman vibrations of the A-T and G-C bases on drug interaction. At r>1/5, a minor helix destabilization occurred with participation of several DNA donor sites in drug complexation. The ascorbate anion interaction occurred mainly through H-bonding of the acid OH and C-O groups with DNA phosphate, bases and doxyribose donor atoms.

Animals↗