Determination of the DNA-binding characteristics of ethidium bromide, proflavine, and cisplatin by flow injection analysis: Usefulness in studies on antitumor drugs

Alonso, Ángel Cabo; Almendral, M.J.; Curto, Y.; Criado, Julio J.; Rodríguez, E.; Manzano, Juan L.

doi:10.1016/j.ab.2006.06.004

Cited by 49 publications

(26 citation statements)

References 12 publications

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“…Another cationic dye EthBr exhibits pronounced binding to nucleic acids, and it is extensively used for DNA staining. In addition, EthBr when bound to DNA exhibits robust increase in fluorescence intensity (1,23,40). Because of these limitations, the exact assessment of the EthBr diffusion rate through the gap junctions is problematic.…”

Section: Resultsmentioning

confidence: 99%

Gap junction permeability: selectivity for anionic and cationic probes

Kanaporis

Brink

Valiūnas

2011

American Journal of Physiology-Cell Physiology

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Gap junction channels formed by different connexins exhibit specific permeability to a variety of larger solutes including second messengers, polypeptides, and small interfering RNAs. Here, we report the permeability of homotypic connexin26 (Cx26), Cx40, Cx43, and Cx45 gap junction channels stably expressed in HeLa cells to solutes with different size and net charge. Channel permeability was determined using simultaneous measurements of junctional conductance and the cell-cell flux of a fluorescent probe. All four connexins allowed passage of both cationic and anionic probes, but the transfer rates were connexin dependent. The negatively charged probes [Lucifer yellow (LY; median axial diameter 9.9 Å, charge -2), carboxyfluorescein (CF; 8.2 Å; -2), and Alexa Fluor350 (AF350, 5.4 Å; -1)] exhibited the following permeability order: Cx43 > Cx45 > Cx26 > Cx40. In contrast, for the positively charged species permeability, the orders were as follows: Cx26 ≈ Cx43 ≈ Cx40 ≈ Cx45 for N,N,N-trimethyl-2-[methyl-(7-nitro-2,1,3-benzoxadiol-4-yl) amino] ethanaminium (NBD-m-TMA; 5.5 Å, +1) and Cx26 ≥ Cx43 ≈ Cx40 > Cx45 for ethidium bromide (10.3 Å, +1). Comparison of probe permeability relative to K(+) revealed that Cx43 and Cx45 exhibited similar permeability for NBD-m-TMA and AF350, indicating weak charge selectivity. However, lesser transfer of CF and LY through Cx45 relative to Cx43 channels suggests stronger size-dependent discrimination of solute. The permeability of NBD-m-TMA for Cx40 and Cx26 channels was approximately three times higher than to anionic AF350 despite the fact that both have similar minor diameters, suggesting charge selectivity. In conclusion, these results confirm that channels formed from individual connexins can discriminate for solutes based on size and charge, suggesting that channel selectivity may be a key factor in cell signaling.

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Section: Resultsmentioning

confidence: 99%

Gap junction permeability: selectivity for anionic and cationic probes

Kanaporis

Brink

Valiūnas

2011

American Journal of Physiology-Cell Physiology

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“…In order to illustrate quantitatively the consequence, changes in emission intensities at about 450 nm for the two Cu(2þ) complexes have been plotted against the added DNA concentration per mole of complexes. According to the Scatchard equation [37][38][39], a plot of r/C f versus r gives the binding constants of complexes 1 and 2. The results imply that both the compounds can insert between DNA base pairs and complex 1 (K ¼ 7.31 Â10 6 ) can interact with DNA more strongly than complex 2 (K ¼ 2.33 Â 10 6 ).…”

Section: Fluorescence Spectramentioning

confidence: 99%

Synthesis, crystal structure, DNA interaction and antioxidant activities of two novel water-soluble Cu(2+) complexes derivated from 2-oxo-quinoline-3-carbaldehyde Schiff-bases

Liu

Wang

Zhou

et al. 2009

European Journal of Medicinal Chemistry

270

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“…The detection of superoxide within the mitochondrial matrix using the TPP-HE probe may be complicated by the uncontrolled enhancement of the OH-TPP-E + and TPP-E + fluorescent responses upon intercalation with mtDNA [15, 17, 25]. As in a previous study in which HE was used as a probe [16], we removed mtDNA in the mitochondrial samples through digestion with DNase 1, thereby successfully eliminating the uncontrolled fluorescence enhancement of OH-TPP-E + and TPP-E + .…”

Section: Methodsmentioning

confidence: 99%

Qualitative determination of superoxide release at both sides of the mitochondrial inner membrane by capillary electrophoretic analysis of the oxidation products of triphenylphosphonium hydroethidine

Arriaga

2009

Free Radical Biology and Medicine

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Superoxide is released asymmetrically to both sides of the mitochondrial inner membrane. Since this membrane is impermeable to superoxide, two separate pools are formed at either side of the membrane, each with its own characteristics and potential biological effects. Here, we report an attomole-sensitive fast capillary electrophoretic method that can analyze superoxide in a single pool, either the matrix pool or that outside the mitochondria. The method uses triphenylphosphonium hydroethidine (TPP-HE) that reacts with the superoxide in both pools. Centrifugation is used to separate the mitochondria (i.e. matrix contents) from the supernatant (i.e. products released outside the mitochondria). Each fraction is then analyzed by a capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) method that separates and detects hydroxytriphenylphosphonium ethidium (OH-TPP-E + ), the fluorescent superoxide specific product. The separation takes < 3 min and the detection level is down to 3 attomole OH-TPP-E + . The method has proved to be effective detecting qualitatively superoxide release in the mitochondria of 143B cells, mouse liver, and rat skeletal muscle, both in the presence and absence of inhibitors. In addition, this study confirmed that Complex I releases superoxide only toward the matrix while Complex III releases superoxide toward both sides of the mitochondrial inner membrane. Furthermore, treatment with menadione induces superoxide release toward both sides of the mitochondrial inner membrane.

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Determination of the DNA-binding characteristics of ethidium bromide, proflavine, and cisplatin by flow injection analysis: Usefulness in studies on antitumor drugs

Cited by 49 publications

References 12 publications

Gap junction permeability: selectivity for anionic and cationic probes

Gap junction permeability: selectivity for anionic and cationic probes

Synthesis, crystal structure, DNA interaction and antioxidant activities of two novel water-soluble Cu(2+) complexes derivated from 2-oxo-quinoline-3-carbaldehyde Schiff-bases

Qualitative determination of superoxide release at both sides of the mitochondrial inner membrane by capillary electrophoretic analysis of the oxidation products of triphenylphosphonium hydroethidine

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