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1.
The photoreaction of the furan-side moiety of 5-methoxypsoralen (5-MOP) with thymidine used as a DNA model compound was investigated in the dry state. Under these conditions, two main fluorescent photoadducts were formed and isolated by HPLC. The two modified nucleosides were characterized as the two cis-syn diastereoisomers of furan-side monoadducts of 5-MOP to thymidine on the basis of spectroscopic measurements including UV, fluorescence, 1H-NMR and circular dichroism analysis. The identification and quantification of the latter photoproducts within naked DNA exposed to photoexcited 5-MOP were achieved by enzymatic digestion completed by HPLC separation and fluorescence detection. Similarly, the two cis-syn furan-side monoadducts were found to be formed in the DNA of Saccharomyces cerevisiae cells after incubation with 5-MOP and subsequent exposure to 365 nm at an incident dose of 38.4 kJ m?2. Under these conditions, the rate of induction of two diastereoisomeric photoadducts was as low as one modification per 106 and 2 × 105 bases, respectively.  相似文献   

2.
Single‐labeled pyridylporphyrin–DNA conjugates are reported as highly sensitive and selective spectroscopic sensors for mercury(II) ions in water. The effects of chemical structure (thymine versus adenine), number of nucleotides (monomer versus octamer), and porphyrin metalation (Zn versus free base) on the sensitivity and selectivity of mercury(II) detection are explored. The results indicated that pyridylporphyrin rather than the nucleobase plays a crucial role in mercury(II) sensing, because porphyrin conjugates with both adenosine and thymidine exhibited excellent mercury(II) detection. Mercury(II) recognition was shown in emission quenching, as well as in a redshift of the porphyrin Soret band absorption. The limit of detection (LOD, 3σ/slope) of zinc(II) pyridylporphyrin‐5′‐oligodeoxythymidine ( ZnPorT8 ) obtained by fluorescence quenching was calculated to be 21.14 nM . Other metal cations (Zn2+, Cd2+, Pb2+, Mn2+, Ca2+, Ni2+, Mg2+, Fe2+, Cu2+, and Na+) did not interfere with the emission and absorption sensing of mercury(II). Free‐base porphyrin–oligothymine conjugate 2HPorT8 displayed similar sensitivity to ZnPorT8 but different selectivity. The results also implied that the sensing properties of porphyrin–deoxythymidine conjugates could potentially be tuned by porphyrin metalation.  相似文献   

3.
To study the interaction between gatifloxacin (GT), metal ions (Cu2+, Cd2+, Co2+, Mg2+) and calf thymus DNA under condition of physiology pH, UV absorption and fluorescence methods were adopted. Result shows that metal ions and DNA are able to react with GT in ground state. In further research, by studying the influence of metal ions on binding of GT with DNA in metal ions–GT–DNA ternary system, we found that influential mechanism of Mg2+ on the binding of GT with DNA may be different from the other three. Mg2+ can act as a bridge in the binding of GT's carboxyl/carbonyl with DNA phosphate in certain concentration range; while Cu2+, Cd2+, Co2+ can combine directly with GT by reaction between GT carboxyl/carbonyl and DNA base, and enhance the binding ability of GT with DNA. The influence extent and type depend not only on the binding site of DNA with metal ions (phosphate or base), but also the binding ability of which. The stronger the binding ability of metal ions with DNA base is, the larger their promotion to binding of GT with DNA is. The order of metal ions’ influential ability on the binding of GT–DNA is identical to the binding ability order of metal ions with DNA base, that is: Cu2+ > Cd2+ > Co2+ > Mg2+.  相似文献   

4.
Abstract— The photomediated induction of interstrand cross-links by 8-methoxypsoralen has been measured in the epidermal DNA of hairless mice. Equivalent efficiencies for cross-link induction were determined for HRS/J/Anl and SKH: hairless-1 mice. A wavelength dependence on the relative efficiency of cross-link induction was observed; a broad spectrum light source, 300–400 nm, was approximately 5 times more effective in cross-link formation than a 365 nm light source. Repeated exposure to 8-methoxypsoralen followed by ultraviolet light, 5 times a week for 6 weeks, altered epidermal thickness and resulted in a decreased efficiency for DNA cross-link formation.  相似文献   

5.
Abstract— We have developed a procedure called a plaque reduction assay to assess the biological activity of duplex circular DNA modified by covalent adduct formation with psoralen derivatives. The replicating form (RF) of bacteriophage DNA modified by photochemical addition of a psoralen derivative was introduced into bacterial cells using the CaCI2 transfection method. The transfected cells. plated upon a confluent lawn of cells permissive for the bacteriophage in the inoculum, provided a measure of the reduction in infectivity of the RF DNA which resulted from its covalent modification. Use of this assay is illustrated in studies which screened and compared the activities of several recently synthesized psoralen derivatives. We describe two new compounds. β-(8-psoralenoxy)-ethanol and β-(8-psoralenoxy)ethylamine that are significantly more active than either 8-methoxypsoralen or trioxsalen in the biological assay  相似文献   

6.
Abstract— The protozoan Tetrahymena thermophila was treated with 8-methoxypsoralen in combination with long wavelength ultraviolet irradiation and the DNA-repair response was studied. Following the treatment a lag-period in cell proliferation was observed, the duration of which was proportional to the amount of psoralen used, and both swelling and deformation of the cells were observed. The treatment with 3 μg 8-methoxypsoralen/mℓ and a light dose of 8 kJ m-2 resulted in a 10-fold decrease in DNA and RNA synthesis activity, while the protein synthesis was only moderately affected. Using the same conditions the lag-period was 30 h, and during this time the psoralen induced DNA interstrand cross-links were removed. Alkaline elution experiments showed that the repair process involves DNA single strand scissions, whereas no double strand DNA scissions were detected.  相似文献   

7.
溶液中N-乙酸基取代氮氧杂大环及其配合物稳定性研究   总被引:1,自引:0,他引:1  
用pH电位滴定法在25℃,0.5mol·L-1KNO3水溶液中测定了三种大环化合物:H2L1(1,12-二氮杂-3,4:9,10-二苯并-5,8-二氧杂环十五烷-N,N'-二乙酸);H3L2(1,12,15-三氮杂-3,4:9,10-二苯并-5,8-二氧杂环十七烷-N,N',N″-三乙酸)和H2L3(1,15-二氮杂-3,4:12,13-二苯并-5,8,11-三氧杂环十八烷-N,N′-二乙酸)的逐级质子化常数.又测定了它们与Cu2+、Ni2+、Pb2+配合物的稳定常数,以及H2L3与镧系金属La3+、Pr3+、Nd3+、Eu3+、Sm3+、Gd3+、Dy3+、Yb3+配合物的稳定常数.讨论了三种大环化合物质子化的一般顺序及其与各种离子配位时稳定性选择规律.说明了影响配位稳定性的有关因素.  相似文献   

8.
Twelve ternary lanthanide complexes RE(sal)3phen (RE3+ = La3+, Pr3+, Nd3+, Sm3+, Eu3+, Gd3+, Tb3+, Dy3+, Ho3+, Tm3+, Yb3+, Lu3+, sal = salicylic acid, phen = phenanthroline) were prepared. Interactions between the complexes and calf thymus DNA (ct‐DNA) were investigated using UV–visible spectrophotometry, fluorescence quench experiment and viscosity measurement. Hypochromicity and red shift of the absorption spectra of complexes were observed in the presence of DNA. The enhanced emission intensity of ethidium bromide (EB) in the presence of DNA was quenched by the addition of lanthanide complexes, which indicated that the lanthanide complexes displaced EB from its binding sites in DNA. Based on the systematic research of the binding constant (Kb) and the fluorescence quenching constant (Kq) of the 12 complexes, we found that the complexes with smaller lanthanide ion radius had stronger binding abilities with DNA. Viscosity measurement showed that the relative viscosity of the DNA solution was enhanced with increasing the amounts of the complexes. All these results suggested that the complexes could bind to DNA and the major binding mode was intercalative binding. Moreover, all these complexes exhibited excellent antibacterial abilities against Escherichia coli. Also, the antibacterial activities of complexes with heavy rare earth were higher than those of complexes with light rare earth. Copyright © 2014 John Wiley & Sons, Ltd.  相似文献   

9.
Several classes of copper complexes are known to induce oxidative DNA damage that mediates cell death. These compounds are potentially useful anticancer agents and detailed investigation can reveal the mode of DNA interaction, binding strength, and type of oxidative lesion formed. We recently reported the development of a DNA electrochemical biosensor employed to quantify the DNA cleavage activity of the well-studied [Cu(phen)2]2+ chemical nuclease. However, to validate the broader compatibility of this sensor for use with more diverse—and biologically compatible—copper complexes, and to probe its use from a drug discovery perspective, analysis involving new compound libraries is required. Here, we report on the DNA binding and quantitative cleavage activity of the [Cu(TPMA)(N,N)]2+ class (where TPMA = tris-2-pyridylmethylamine) using a DNA electrochemical biosensor. TPMA is a tripodal copper caging ligand, while N,N represents a bidentate planar phenanthrene ligand capable of enhancing DNA interactions through intercalation. All complexes exhibited electroactivity and interact with DNA through partial (or semi-) intercalation but predominantly through electrostatic attraction. Although TPMA provides excellent solution stability, the bulky ligand enforces a non-planar geometry on the complex, which sterically impedes full interaction. [Cu(TPMA)(phen)]2+ and [Cu(TPMA)(DPQ)]2+ cleaved 39% and 48% of the DNA strands from the biosensor surface, respectively, while complexes [Cu(TPMA)(bipy)]2+ and [Cu(TPMA)(PD)]2+ exhibit comparatively moderate nuclease efficacy (ca. 26%). Comparing the nuclease activities of [Cu(TPMA)(phen)] 2+ and [Cu(phen)2]2+ (ca. 23%) confirms the presence of TPMA significantly enhances chemical nuclease activity. Therefore, the use of this DNA electrochemical biosensor is compatible with copper(II) polypyridyl complexes and reveals TPMA complexes as a promising class of DNA damaging agent with tuneable activity due to coordinated ancillary phenanthrene ligands.  相似文献   

10.
6-苄氨基嘌呤及其金属配合物与DNA的作用机理   总被引:4,自引:0,他引:4  
席小莉  杨曼曼  杨频 《无机化学学报》2005,21(12):1847-1852
新型抗癌症药剂的开发和抗癌机理研究,一直是人们争相研究的前沿课题[1-5]。而许多预防和治疗癌症的药物都是以D N A为作用靶来设计的[6]。以D N A为靶分子的金属抗癌剂,其抗癌活性是由于金属离子与D N A的配位,即金属离子与D N A的某些亲核基团(如磷酸氧位点或碱基氮、氧位点)  相似文献   

11.
Studies of double‐stranded‐DNA binding have been performed with three isomeric bis(2‐(n‐pyridyl)‐1H‐benzimidazole)s (n=2, 3, 4). Like the well‐known Hoechst 33258, which is a bisbenzimidazole compound, these three isomers bind to the minor groove of duplex DNA. DNA binding by the three isomers was investigated in the presence of the divalent metal ions Mg2+, Co2+, Ni2+, Cu2+, and Zn2+. Ligand–DNA interactions were probed with fluorescence and circular dichroism spectroscopy. These studies revealed that the binding of the 2‐pyridyl derivative to DNA is dramatically reduced in the presence of Co2+, Ni2+, and Cu2+ ions and is abolished completely at a ligand/metal‐cation ratio of 1:1. Control experiments done with the isomeric 3‐ and 4‐pyridyl derivatives showed that their binding to DNA is unaffected by the aforementioned transition‐metal ions. The ability of 2‐(2‐pyridyl)benzimidazole to chelate metal ions and the conformational changes of the ligand associated with ion chelation probably led to such unusual binding results for the ortho isomer. The addition of ethylenediaminetetraacetic acid (EDTA) reversed the effects completely.  相似文献   

12.
In this article, we describe the analysis of aptamers for Hg2+ ions through CE with LIF (CE‐LIF) detection using 2% poly(ethylene oxide) solutions containing OliGreen (fluorophore). In the presence of an EOF, DNA strands migrating against the EOF were detected at the cathode end. Four DNA strands – T33, T5C28, T5C5T23, and T15C5T13 – could not be separated through CE‐LIF in the absence of Hg2+. At 0.3 mM Hg2+, however, all four were partially separated within 20 min, with SDs of the migration times all being less than 2.5%. From the CE, fluorescence, and ellipticity data, we concluded that the conformations of these four DNA strands all changed from random‐coil to folded structures as a result of T–Hg2+–T bonding. In addition, we found that this CE approach provided different electropherograms patterns for T7, T15, and T33 in the absence and presence of Hg2+, indicating various interactions of the DNA strands with Hg2+. Using this simple, high‐resolution CE approach, we also demonstrated that adenosine triphosphate has a stronger interaction with the adenosine triphosphate aptamer than with either the platelet‐derived growth factor aptamer or T33. This CE approach holds great potential for screening aptamers for small solutes, studying the catalytic activity of DNAzymes, and evaluating the biological functions of microRNA.  相似文献   

13.
Abstract— Isolated calf thymus DNA was treated with the 1,2-dioxetanes 3-acetoxymethyl-3,4,4-tri-methyl-1,2-dioxetane, 2,3-dimethylbenzofuran dioxetane, 3-hydroxymethyl-3,4,4-trimethyl-1,2-dioxeta-ne (HTMD), 3,3,4,4-tetramethyl-1,2-dioxetane and 3,4,4-trimethyl-1,2-dioxetane (TrMD), which on thermal decomposition generate triplet-excited carbonyl products. To monitor quantitatively the formation of the mutagenic oxidation product 7,8-dihydro-8-oxoguanine (8-oxoGua), a sensitive and selective HPLC electrochemical assay was used after acidic hydrolysis (HF/pyridine) of the dioxetane-treated DNA. High yields of 8-oxoGua (up to ca 4% of the available guanine) were obtained for HTMD and TrMD. Both were investigated in detail with respect to effects of concentration, time and temperature. The oxidative reactivity of 1,2-dioxetanes was compared with several type I (benzophenone and riboflavin) and type II (methylene blue and rose bengal) photooxidants and disodium 1,4-etheno-2,3-ben-zodioxin-1,4-dipropionate as a chemical source of singlet oxygen. The persistence of 8-oxoGua towards oxidation by HTMD was examined in the reaction with 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxodGuo) and with oxidized DNA. It was shown that, indeed, 8-oxoGua is consumed in the oxidized DNA on prolonged exposure to an excess of HTMD. The reaction of 8-oxodGuo with HTMD afforded the two 4R* and 4S* diastereomers of 9-(2-deoxy-ß-D-erythropentofuranosyl)-4,8-dihydro-4-hydroxy-8-oxoguanine as main oxidation products. Trapping experiments with teft-butanol confirmed that hydroxyl radicals are not involved, whereas the use of the triplet quenchers sodium 9,10-dibromo-anthra-cene-2-sulfonate and 2,3-diazabicyclo[2.2.1]hept-2-ene established that triplet-excited states are mainly responsible for the observed DNA oxidation through type I action (electron transfer chemistry). The role of singlet oxygen was tested by means of deuterium isotope effects in D2O versus H2O, but no definitive conclusion could be reached in regard to the involvement of 102 in these oxidations. The present results reveal that 1,2-dioxetanes are efficient DNA oxidants and excellent tools to study photooxidation reactions of DNA in the dark.  相似文献   

14.
Abstract— In Saccharomyces cerevisiae, a re-irradiation with a saturing dose of UVA after pretreatment with 8-methoxypsoralen (8-MOP) plus low doses of UVA and removal of unbound 8-MOP lead to an increase in frequency of forward mutants in strains defective in the excision (radl-3, radl-Δ, rad2-6) or in the recombinational (rad52-l) repair pathways. Such an enhancement attributable to DNA interstrand cross-links was not observed in mutants blocked in a mutagenic repair pathway (rad6-Δ and pso2-l). These results are interpreted as revealing the existence of an alternative pathway to excision of DNA cross-links. This pathway appears to be error-prone and independent from the recombinational pathway. The RAD6 or the PSO2 gene products are likely to interfere with this process.  相似文献   

15.
Abstract— The proportions of single-strand breaks and alkali-labile bonds produced by UV-light were investigated in covalently-closed circular 5-bromouracil (BrUra)-containing λ-phage DNA. When BrUra DNA was irradiated in 001 M Tris-0–001 M EDTA (pH 8-1) buffer, the D0 was 11-7 J/m2 for single-strand breaks, 2–25 J/m2 for total breaks, and 2–8 J/m2 for alkali-labile bonds. Thus, alkali-labile bonds were the predominant photochemical products. No double-strand breaks were observed after exposure to 7-7 times the D0 for neutral breakage. The photolability measured under both neutral and alkaline conditions was affected by the NaCl concentration in the irradiation solvent, with the greatest resistance to breakage exhibited at the lowest concentrations. The composition of the irradiation buffer also affected sensitivity. Exposure in 1/10 SSC yielded 4-4 (neutral) and 5–7 (alkaline) times the breakage produced in Tris-EDTA.  相似文献   

16.
The quantum yield of singlet oxygen generation by aqueous furocoumarins was measured at 365 nm using the photosensitized inactivation of subtilisin Carlsberg as the probe with the following results: psoralen (0.18), 5-methoxypsoralen (0.013), and 8-methoxypsoralen (0.035). Singlet oxygen formation was significant for dark complexes of 8-MOP with calf thymus DNA and the covalent DNA photoadducts. Incorporation of 8-MOP in sonicated egg phosphatidylcholine liposomes did not inhibit photosensitization of subtilisin Carlsberg and also led to lipid peroxidation, with positive tests for the involvement of singlet oxygen. Peroxidation of the liposomes was inhibited by the presence of α-tocopherol and promoted by the presence of cholesterol in the membranes.  相似文献   

17.
Abstract— Calf thymus DNA was irradiated with low-intensity UVA (main output at 365 nm, 2 mW cm?2 or 36 kj m 2 for 30 min), and the role of metal ions, hydrogen peroxide and reactive oxygen species (ROS) was examined. DNA damage was measured as thiobarbituric acid-reactive substances (possibly from degradation of deoxyribose) and as changes in ethidium bromide-DNA fluorescence due to unwinding from strand breaks. Under the present experimental conditions, UVA alone or in the presence of H202 had no effect on DNA but slightly enhanced the damage by iron/EDTA. Ultraviolet A strongly enhanced DNA damage (ca four- to five-fold) by the Fenton reaction system (50 μM Fe2+/100 μM EDTA + 0.5 mM H202). The results suggest that the Fenton reaction system was “photosensitized” to damage DNA by low-intensity UVA radiation. The enhanced damage by UVA was attributed in part to the reduction of Fe3+ to Fe2+. Ultraviolet A had no effect when iron (ferric or ferrous) ions were replaced by Cu2+, Zn2+, Mn2+ or Cd2+. The ROS involved in the UVA-enhanced damage to DNA by the Fenton reagents were OH and, to a lesser extent, superoxide anions. The UVA-potentiated DNA damage by the Fenton reaction system was then used to examine the protective effect of para-aminobenzoate (PABA), a UVB-absorbing sunscreen that protects against photocarcinogenesis in hairless mice. The results show that PABA and mannitol dose-dependently inhibited the damage with concentrations required for 50% inhibition at 0.1 mM and 3 mM, respectively. The protection by PABA was attributed to its radical-scavenging ability because PABA does not absorb light in the UVA region. These findings may be relevant to the biological damage by UVA and suggest that PABA is useful in protection against photocarcinogenesis by wide-range UV radiation.  相似文献   

18.
Seven new neutral mononuclear metal complexes of VO2+, Mn2+, Fe3+, Co2+, Ni2+, Zn2+ and Cd2+ with the quinolone antibacterial agent oxolinic acid (=Hoxo) have been prepared and characterized with physicochemical and spectroscopic techniques. In all the complexes, oxolinic acid acts as a bidentate deprotonated ligand bound to the metal through the pyridone oxygen and one carboxylate oxygen. The metals in all the complexes are six-coordinate with slightly distorted octahedral geometry. The lowest energy model structures of the complexes Fe(oxo)3, VO(oxo)2(H2O) and Mn(oxo)2(H2O)2 have been determined with molecular modeling calculations. The ability of all the complexes to bind to calf-thymus DNA has been investigated with diverse spectroscopic techniques.  相似文献   

19.
Graphene quantum dots (GQDs), inheriting the superb property of graphene oxide, possess smaller lateral size and high biocompatibility, thus having potential in biomedical applications. We previously discovered that GQDs, combining with Cu2+ ions, could cleave DNA primarily through an oxidative pathway; yet, oxidative DNA cleavage is not practically preferred in biology. In this work, we explore the DNA cleavage ability of GQDs with Zn2+ and Ni2+. Zn2+ and Ni2+ alone are incapable of cleaving supercoiled DNA, but when combining with the GQDs, Zn2+ and Ni2+ exhibit DNA cleavage activity. However, the activity of these two systems is much lower than that of GQDs/Cu2+, and GQDs/Ni2+ is less active than GQDs/Zn2+. The functional mechanism of GQDs/Ni2+ and GQDs/Zn2+ is different from that of GQDs/Cu2+. The GQDs play a key role in the two systems; the redox inactive Zn2+ and Ni2+ ions assist to generate the oxidative species that eventually lead to the DNA cleavage. The current results together with our previous result indicate that GQDs together with metal ions can cleave supercoiled DNA, and their cleavage activities depend on the properties of metal ions: for redox active metal ions, metal ions play key roles, for redox inactive metal ions, GQDs are dominant.  相似文献   

20.
Three series of metal salophen complexes derived from Zn2+, Cu2+, Pt2+ and Ni2+ have been synthesized and their interaction with quadruplex DNA has been evaluated. The compounds differ on the number of ethyl piperidine substituents. They have been characterized by 1H NMR, IR and UV-visible spectroscopies and by HR-mass spectrometry. Their luminescent properties have been also evaluated and we can observe that, as expected, Zn2+ and Pt2+ complexes are those displaying more interesting luminescence with an emission band red-shifted with respect to the corresponding uncoordinated ligand. DNA interactions with G4 and duplex DNA were evaluated by FRET melting assays (for the Zn2+, Cu2+ and Ni2+ complexes) and by emission titrations (for one Pt2+ complex) which indicated that the disubstituted compounds 2-Ni and 2-Pt are the only ones that display good affinity for G4 DNA structures.  相似文献   

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