ABSTRACT
Absorption and fluorescence emission properties of an N-salicylideneamine fluorescent dye molecule N, N'-bis(2-hydroxy-5-methylbenzylidene)-1,2-ethanediamine (1) have been studied in three typical solvents--2-methylbutane, ethanol, dimethyl sulphoxide (DMSO), and its DNA complex in methanol/H2O mixed solvent. The normal absorption band of 1 is observed in both aprotic and protic solvents and has been assigned to the l --> a(pi) transition in the enol form of 1. The long-wavelength absorption band of 1, which is caused by the formation of a cis-keto species in the ground state, is absent in aprotic solvents, but is observable in protic ones. Normal fluorescence emission from the excited enol state of 1 is obtained only when the normal absorption band is excited, while the excited-state intramolecular proton transfer (ESIPT) emissions from both cis- and trans-keto species are recorded in all cases, being acceptable for the variation of the relative emission intensities. A preliminary spectroscopic study of the 1-DNA complex indicates an intercalation-binding mode, the convincing supporting evidence being the enhanced ESIPT fluorescence intensity of 1 when complexed with DNA. Finally, a universal energy-state diagram is given to interpret the experimental results.
Subject(s)
DNA/chemistry , Fluorescent Dyes/chemistry , Animals , Benzylamines/chemistry , In Vitro Techniques , Salicylates/chemistry , Solutions , Solvents , Spectrometry, FluorescenceABSTRACT
Methylene blue (MB) is a phenothiazinium photosensitizer with promising applications in the photodynamic therapy (PDT) for anticancer treatment. The binding properties of MB to herring sperm DNA have been investigated by the measurements of absorption spectra, quenching experiments and the elucidation of the photobleaching processes. Remarkable hypochromic and bathochromic effects of MB in the presence of increasing amounts of DNA have been observed in the absorption spectra. The quenching of MB by the DNA bases obeys the Stern-Volmer equation and ferrocyanide quenching of MB in the absence and presence of DNA is also measured as extended experiments. Results from the above spectral measurements are all consistent with the intercalative binding mode of MB to DNA with the Kb value of 1.89 x 10(4) M(-1). The photobleaching processes of MB and its DNA complex have also been studies, which indicate that the photobleaching of MB and its DNA complex proceeds with different mechanisms and the reactive oxygen species are responsible for the self-sensitized photooxidation of MB.
Subject(s)
DNA/chemistry , Fishes/metabolism , Methylene Blue/chemistry , Photosensitizing Agents/chemistry , Spermatozoa/metabolism , Animals , DNA/metabolism , Ferrocyanides/chemistry , Male , Methylene Blue/metabolism , Molecular Structure , Photobleaching/radiation effects , Photosensitizing Agents/metabolism , Spectrum Analysis , Spermatozoa/radiation effectsABSTRACT
Two inhibitor-containing 'half-sandwich' cobalt(II) complexes [(TpPh)Co(X)(CH3OH)m] x nCH3OH ((TpPh) = hydrotris (3-phenylpyrazolyl)borate; 1: X- = N3-, m = 1, n = 2; 2: X- = NCS-, m = 0, n = 0) have been synthesized and used as the catalysts in the bicarbonate dehydration reaction. The structures of 1 and 2 were determined by X-ray diffraction analysis, which shows that N3- and NCS- coordinate to the Co(II) ions of 1 and 2, respectively, with the Co-N bond lengths of 1.992(6) A and 1.901(3) A. The coordination geometries of the Co(II) complexes in solution are five-coordinated trigonal bipyramid as revealed by the spectroscopic measurements. The dehydration kinetic measurements of HCO3- are performed by the stopped-flow techniques at pH < 7.9. The apparent dehydration rate constant k(obs) varies linearly with Co(II) complex and H+ concentrations, respectively, and the catalytic activity of 2 is lower than that of 1. The aqua Co(II) complex must be the reactive catalytic species in the catalyzed dehydration reaction and the rate-determining step is the substitution of the labile water molecule by HCO3-. The k(obs) values increase with increasing reaction temperature, and the large negative entropy of activation also indicates the associative activation mode. The inhibition ability of NCS- is stronger than that of N3-, which can be rationalized by the decreases in the Co-N(N3-/NCS-) bond lengths and effective atomic charges of the Co(II) ions based on the X-ray crystallographic data and theoretical calculations in this work.
Subject(s)
Bicarbonates/chemistry , Carbon Dioxide/chemistry , Organometallic Compounds/chemistry , Water/chemistry , Azides/chemistry , Binding Sites , Borates/chemistry , Carbonic Anhydrase Inhibitors , Carbonic Anhydrases , Catalysis , Cobalt/chemistry , Crystallography, X-Ray , Kinetics , Molecular Conformation , Organometallic Compounds/chemical synthesis , Spectrophotometry, Infrared , Spectrophotometry, Ultraviolet , Thiocyanates/chemistryABSTRACT
Spectroscopic studies of the interactions of Ni(II) ion with herring sperm DNA have been performed in this contribution using methylene blue (MB) as the fluorescent probe molecule. MB binds to double helical DNA via the intercalative mode, and its fluorescence is efficiently quenched by the DNA nucleobases. The fluorescence intensity of the probe molecule increases distinctly when Ni(II) ion is added to the MB-DNA solution system. These results indicate that the Ni(II) ions may be complexed with the DNA helix and probably bind at N-7 of adenine or guanine; consequently, some intercalated MB molecules are released due to the binding of Ni(II) ions to N-7 nitrogen of the purines. The Ni(II) ion-DNA interactions are further investigated by performing the photobleaching experiments of the MB-DNA complex in the presence and absence of Ni(II) ion. The bi-exponential decay functions of the fluorescence intensity have been observed in both cases and the shortening of the slow decay component when added Ni(II) ion also agrees with the release of MB from the DNA duplex.
Subject(s)
DNA/chemistry , Fluorescent Dyes/chemistry , Methylene Blue/chemistry , Nickel/chemistry , Adenine/chemistry , Animals , Fishes , Guanine/chemistry , Molecular Structure , Nitrogen/chemistry , Photobleaching , Spectrometry, FluorescenceABSTRACT
A series of half-sandwich copper(II) complexes [TpPh]CuX ([TpPh] = hydrotris(3-phenyl-pyrazolyl)borate; X- = OH- (1), N3- (2), NCS- (3)) have been synthesized as models for carbonic anhydrase. The structure of 3 was determined by X-ray diffraction analysis. Crystals of 3 (C37H30BCuN9S) are triclinic, space group P1 with a = 11.997(3) A, b = 12.116(3) A, c = 13.384(4) A, alpha = 81.088(5) degrees, beta = 79.289(6) degrees, gamma = 68.668(5) degrees, V = 1772.4(8) A3, and Z = 2. The dehydration kinetic measurements of HCO3- are performed by the stopped-flow techniques at pH < 7.9. The apparent dehydration rate constant kdobs varies linearly with total Cu(II) concentration, and the catalytic activity of the model complexes decreases in the order 1 > 2 > 3. The catalytic activity decreases with increasing pH indicating that the aqua model complex must be the reactive catalytic species in the catalyzed dehydration reaction and the rate-determining step is the substitution of the labile water molecule by HCO3-. The kdobs values increase with increasing reaction temperature, and the apparent activation energies of the model complexes with inhibitors are remarkably higher than that of the complex with no inhibitors, this being the origin of inhibition. The large negative entropy of activation also indicates an associative mode of activation in the rate-determining step. The inhibition ability of the inhibitor NCS- is stronger than that of the inhibitor N3-, which can be rationalized by the decrease in effective atomic charges of the Cu(II) ions as revealed by the theoretical calculations.
