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1.
J Inorg Biochem ; 77(1-2): 43-6, 1999 Oct.
Article in English | MEDLINE | ID: mdl-10626352

ABSTRACT

In this work, the B-->Z transition of poly(dG-dC).poly(dG-dC) and the B-->A transition of poly(dG).poly(dC) and of calf thymus (CT) DNA fragments modified by antitumor bifunctional polynuclear platinum complexes were investigated by circular dichroism (CD). The transition from the B- to Z-form of DNA was inducible with all three compounds studied, as indicated by an inversion of the B-form spectra. The B-->A transition in poly(dG).poly(dC) was induced easily by platinum complex binding alone, while the B-->A transition in CT DNA was induced by ethanol but inhibited by coordination of all polynuclear platinum compounds used here. It was shown that the compound [¿cis-PtCl(NH3)2¿2 mu-¿H2N(CH2)6NH2¿] (NO3)2 (1,1/c,c) was most effective at inhibiting the B-->A transition in CT DNA, and [¿trans-PtCl(NH3)2¿2 mu-¿trans-Pt(NH3)2(H2N(CH2)6NH2)2¿] (NO3)4 (1,0,1/t,t,t) was least effective, while the effectiveness of [¿trans-PtCl(NH3)2¿2 mu-¿H2N(CH2)6NH2¿] (NO3)2 (1,1/t,t) fell between the two. This corresponded to the relative amounts of interstrand crosslinks in double-stranded DNA caused by each compound.


Subject(s)
Antineoplastic Agents/pharmacology , DNA/chemistry , DNA/drug effects , Organoplatinum Compounds/pharmacology , Animals , Antineoplastic Agents/metabolism , Cattle , Circular Dichroism , Cisplatin/chemistry , Cisplatin/pharmacology , Cross-Linking Reagents/chemistry , Cross-Linking Reagents/pharmacology , DNA/metabolism , Dose-Response Relationship, Drug , Nucleic Acid Conformation/drug effects , Organoplatinum Compounds/chemistry , Organoplatinum Compounds/metabolism , Poly C/chemistry , Poly G/chemistry , Polydeoxyribonucleotides/chemistry , Structure-Activity Relationship , Substrate Specificity
2.
Eur J Biochem ; 254(3): 547-57, 1998 Jun 15.
Article in English | MEDLINE | ID: mdl-9688265

ABSTRACT

Recent observations that several trans-platinum complexes exhibit antitumor activity including activity in cisplatin-resistant tumor cells, violates the classical structure/activity relationships of platinum(II) complexes. According to these relationships, only bifunctional platinum(II) complexes with cis-oriented leaving ligands should be therapeutically active. In order to contribute to the understanding of mechanisms underlying the antitumor activity of these new trans-platinum analogs, various biochemical and biophysical methods as well as molecular modeling techniques were employed to study the modifications of DNA by antitumor trans-[PtCl2(NH3)(quinoline)]. The results indicated that trans-[PtCl2(NH3)(quinoline)] coordinated monofunctionally to DNA with a similar rate as transplatin. The overall rate of the rearrangement to bifunctional adducts was also similar to that observed in the case of DNA modification by transplatin, i.e. it was relatively slow (after 48 h approximately 34% adducts remained monofunctional). In contrast to transplatin, however, trans-[PtCl2(NH3)(quinoline)] formed considerably more interstrand cross-links after 48 h (approximately 30%) with a much shorter half-time (approximately 5 h) (approximately 12% for transplatin, t1/2 > 11 h). The results also suggested that the quinoline ligand in all or in a significant fraction of DNA adducts of trans-[PtCl2(NH3)(quinoline)], in which platinum is coordinated to base residues, was well positioned to interact with the duplex. The adducts of trans-[PtCl2(NH3)(quinoline)] terminated in vitro RNA synthesis preferentially at guanine residues. Surprisingly, the type and extent of conformational alterations induced in DNA indicates that trans-[PtCl2(NH3)(quinoline)] behaves in some respects like cisplatin, as indicated by the fact that trans-[PtCl2(NH3)(quinoline)]-modified DNA is recognized by cisplatin-specific antibodies. Models for both monofunctional adducts and bifunctional interstrand cross-links are proposed. Computer-generated AMBER models show that the combination of monofunctional covalent binding and a stacking interaction between quinoline and the DNA bases can produce a kink in the duplex which is strongly suggestive of the directed bend produced by the major cisplatin-DNA adduct (1,2 intrastrand cross-link). Unique DNA adducts of this type formed by trans-[PtCl2(NH3)(quinoline)] may contribute to the antitumor efficacy of this agent.


Subject(s)
Antineoplastic Agents/pharmacology , DNA/drug effects , Organometallic Compounds/pharmacology , Organoplatinum Compounds , Animals , Antineoplastic Agents/metabolism , Base Sequence , Cattle , DNA/chemistry , DNA/metabolism , DNA Adducts , Ethidium/chemistry , Fluorescent Dyes/chemistry , Molecular Sequence Data , Organometallic Compounds/metabolism , Thiourea/chemistry
3.
Mol Pharmacol ; 53(5): 846-55, 1998 May.
Article in English | MEDLINE | ID: mdl-9584210

ABSTRACT

We synthesized a novel platinum drug, cis-[PtCl(NH3)2(N7-ACV)]+, in which ACV is the antiviral drug acyclovir [a deoxyriboguanosine analogue, 9-(2-hydroxyethoxymethyl)guanine]. This new compound exhibits antiviral efficacy in vitro and exhibits an antitumor activity profile different from that of cisplatin [Metal-Based Drugs 2:249-256 (1995)]. To contribute to understanding the mechanisms underlying biological activity of this new compound, we studied modifications of natural and synthetic DNAs in cell-free media by cis-[PtCl(NH3)2(N7-ACV)]+ by various biochemical and biophysical methods. The results indicated that the major DNA adduct of cis-[PtCl(NH3)2(N7-ACV)]+ was a stable monofunctional adduct at guanine residues. In contrast to DNA adducts of other monodentate and clinically ineffective platinum(II) compounds, the adducts of cis-[PtCl(NH3)2(N7-ACV)]+ terminated in vitro DNA and RNA synthesis. In addition, although DNA adducts of cis-[PtCl(NH3)2(N7-ACV)]+ and cisplatin were different, some properties of DNA modified by either compound were qualitatively similar. Such similarities were not noticed if DNA modifications by other ineffective monofunctional platinum(II) complexes were investigated. Thus, the DNA binding mode of monofunctional cis-[PtCl(NH3)2(N7-ACV)]+ was different from that of other monofunctional but ineffective platinum(II) complexes. It has been suggested that the unique capability of cis-[PtCl(NH3)2(N7-ACV)]+ to modify DNA may be relevant to a distinct antitumor efficiency of this novel drug in comparison with cisplatin. It also has been suggested that at least some aspects of DNA interactions of cis-[PtCl(NH3)2(ACV)]+ revealed in the current study could be exploited in the search for and development of new antiviral platinum complexes containing, as a part of the coordination sphere, antiviral nucleosides.


Subject(s)
Acyclovir/analogs & derivatives , Antineoplastic Agents/pharmacology , DNA/drug effects , Organoplatinum Compounds/pharmacology , Acyclovir/pharmacology , Animals , Base Sequence , Cattle , Molecular Sequence Data , Spectrometry, Fluorescence
4.
Mol Pharmacol ; 52(3): 354-61, 1997 Sep.
Article in English | MEDLINE | ID: mdl-9281596

ABSTRACT

Recent findings that an analogue of clinically ineffective transplatin, trans-[PtCl2(E-iminoether)2], exhibits antitumor activity has helped reevaluation of the empirical structure-antitumor activity relationship generally accepted for platinum(II) complexes. According to this relationship, only the cis geometry of leaving ligands in the bifunctional platinum(II) complexes, should be therapeutically active. Global modifications of natural DNAs in cell-free media by trans-[PtCl2(E-iminoether)2] were studied through various molecular biophysical methods and compared with modifications by cis-[PtCl2(E-iminoether)2], transplatin, cisplatin, and monofunctional chlorodiethylenetriamineplatinum(II) chloride. Thus, the results of this study have extended our recent finding, indicating that the prevalent lesion occurring in double-helical DNA on its modification by trans-[PtCl2(E-iminoether)2] is a monofunctional adduct at guanine residues. The modification by trans-[PtCl2(E-iminoether)2] has been found to induce local distortions in DNA, which have a character differing fundamentally from those induced by both clinically ineffective or antitumor platinum complexes tested in this study. The different character of alterations induced in DNA by the adducts of trans-[PtCl2(E-iminoether)2] and transplatin has been suggested to be relevant to the unexpected observation that the new complex with leaving chloride groups in trans position exhibits antitumor efficacy. In addition, the results support the idea that platinum drugs that bind to DNA in a manner fundamentally different from that of cisplatin can exhibit altered biological properties, including differing spectra and intensities of antitumor activity.


Subject(s)
Antineoplastic Agents/pharmacology , DNA/drug effects , Nucleic Acid Conformation/drug effects , Organometallic Compounds/pharmacology , Animals , Cattle , Chemical Phenomena , Chemistry, Physical , Cisplatin/pharmacology , DNA/chemistry , DNA/metabolism , DNA, Superhelical/drug effects , DNA, Superhelical/metabolism , Drug Stability , Stereoisomerism , Substrate Specificity , Transcription, Genetic/drug effects
5.
Eur J Biochem ; 246(2): 508-17, 1997 Jun 01.
Article in English | MEDLINE | ID: mdl-9208945

ABSTRACT

Modifications of natural DNA in a cell-free medium by dinuclear bisplatinum complexes with equivalent coordination spheres, represented by the general formula [¿trans-PtCl(NH3)2¿2(H2N-R-NH2)]2+, where R is a propane or hexane, were studied by various methods of biochemical analysis or molecular biophysics. These methods include binding studies by means of differential-pulse polarography, measurements of melting curves with the aid of absorption spectrophotometry, measurements of CD spectra, ELISA with specific antibodies that recognize DNA modified by platinum complexes, interstrand cross-linking assay employing gel electrophoresis under denaturing conditions and mapping of DNA adducts by means of transcription assays. The results indicated that the major adduct of [¿trans-PtCl(NH3)2¿2(H2N-R-NH2)]2+ in DNA was an interstrand cross-link which was formed with a relatively short half-time (approximately 1 h). At least some types of these interstrand cross-links induced local denaturational changes in the DNA. The results of analyses of interactions of [¿trans-PtCl(NH3)2¿2(H2N-R-NH2)]2+ with linear DNA at relatively higher levels of the modification could be interpreted to mean that these dinuclear platinum complexes were also capable of intrastrand-cross-link formation between adjacent base residues in DNA. However, these intrastrand adducts of [¿trans-PtCl(NH3)2¿2(H2N-R-NH2)]2+ distorted DNA conformation in a way different from the DNA intrastrand adducts of cisplatin. In addition, the DNA adducts of the dinuclear platinum complexes inhibited DNA transcription in vitro. The length of the aliphatic linker chain affected the DNA-binding mode of [¿trans-PtCl(NH3)2¿2(H2N-R-NH2)]2+ and the resulting conformational changes in DNA. The extensive analysis of DNA interactions with [¿trans-PtCl(NH3)2¿2(H2N-R-NH2)]2+ described in this communication has provided further experimental support for previous suggestions [Farrell, N. (1991) in Platinum and other metal coordination compounds in cancer chemotherapy (Howell, S. B., ed.) pp. 81-91, Plenum Press, New York] that the binding of the dinuclear platinum complexes modifies DNA in a way that is different from the modification by antitumor cisplatin. Thus, the results of this work are consistent with the hypothesis that platinum drugs that bind to DNA in a manner fundamentally different from that of cisplatin can exhibit altered biological properties, including a different spectrum and intensity of antitumor activity.


Subject(s)
Antineoplastic Agents/chemistry , Cisplatin/chemistry , DNA/chemistry , Animals , Bacteriophage T7/metabolism , Base Sequence , Cattle , Circular Dichroism , Cross-Linking Reagents/chemistry , DNA/genetics , DNA/metabolism , DNA-Directed RNA Polymerases/metabolism , Immunochemistry , Molecular Sequence Data , Transcription, Genetic , Viral Proteins
6.
Biochemistry ; 33(6): 1316-22, 1994 Feb 15.
Article in English | MEDLINE | ID: mdl-8312249

ABSTRACT

The effects on the conformation of DNA produced by the monofunctional adducts of chloro-(diethylenetriamine)platinum(II) chloride or cis-diamminemonoaquamonochloroplatinum(II) have been investigated by means of the single-strand-specific probe chloroacetaldehyde (CAA). The denatured sites to which CAA was bound and that were induced in DNA by the monofunctional adducts of the platinum complexes were characterized by means of three experimental approaches. These include measurement of the fluorescence of a plasmid fragment treated with CAA, analysis of oligonucleotides treated with CAA and cleaved by piperidine, and termination of duplex transcription on a fragment of plasmid DNA treated with CAA. The results indicate that the denaturational change preferentially occurs in the base pair containing the monoadducted deoxyriboguanosine in the trinucleotide sequence Py-deoxyriboguanosine-Py (Py is a pyrimidine deoxyribonucleoside). It was suggested that this conformational alteration facilitates in DNA the formation of minor bifunctional adducts of cis-diamminedichloroplatinum(II).


Subject(s)
Cisplatin/analogs & derivatives , DNA/chemistry , Nucleic Acid Conformation/drug effects , Nucleic Acid Denaturation/drug effects , Acetaldehyde/analogs & derivatives , Acetaldehyde/metabolism , Base Composition , Base Sequence , Chromosome Mapping , Cisplatin/pharmacology , DNA/metabolism , Deoxyribonucleases, Type II Site-Specific , Enzyme-Linked Immunosorbent Assay , Hydrogen Bonding , Molecular Sequence Data , Piperidines/metabolism , Spectrometry, Fluorescence , Transcription, Genetic
7.
Eur J Biochem ; 216(1): 183-7, 1993 Aug 15.
Article in English | MEDLINE | ID: mdl-8365404

ABSTRACT

The effect of binding of cis-diamminedichloroplatinum(II), its trans isomer and diethylenetriaminechloroplatinum(II) chloride to DNA on the splicing effectiveness of BamHI, EcoRI and SalI restriction endonucleases has been determined by means of gel electrophoresis. All three platinum complexes inhibit the cleavage of linearized plasmid DNA. In addition, the three platinum complexes bound to DNA constitute a barrier across which the linear diffusion of EcoRI on DNA is difficult. We interpret these findings to mean that the splicing effectiveness of restriction enzymes is influenced by bifunctional and monofunctional DNA adducts of platinum via both steric interference and DNA conformational distortions. Whereas the platinum adducts in the restriction sites or in their very close proximity inhibit the cleavage, the lesions occurring a greater distance from the restriction site can slow down the process of the localization of recognition sequences.


Subject(s)
Cisplatin/pharmacology , DNA/metabolism , Deoxyribonucleases, Type II Site-Specific/metabolism , Organoplatinum Compounds/pharmacology , Binding Sites , Deoxyribonuclease BamHI/metabolism , Deoxyribonuclease EcoRI/metabolism , Electrophoresis, Agar Gel , Plasmids/drug effects , Stereoisomerism
8.
Gen Physiol Biophys ; 11(6): 579-88, 1992 Dec.
Article in English | MEDLINE | ID: mdl-1338050

ABSTRACT

The effect of binding of an antitumour drug cis-diamminedichloroplatinum(II) (cis-[Pt(NH3)2Cl2]) to DNA on cutting effectiveness of BamHI, EcoRI, and SalI restriction endonucleases was quantitatively determined. The platinum complex inhibits the cleavage of plasmid pHC624 DNA linearized by BglI restrictase. From the present results we conclude that the yield of restriction endonuclease cleavage is also lowered if the platinum complex is bound outside the recognition DNA sequence of these enzymes. We propose that the origin of platinum adducts on DNA outside the recognition sequence can decrease the yield of restriction enzyme cleavage via inducing a conformational perturbation in the recognition DNA sequence of these enzymes and also via inhibition of the linear diffusion of these enzymes on DNA.


Subject(s)
Cisplatin/pharmacology , DNA/drug effects , Base Sequence , Binding Sites , Cisplatin/chemistry , DNA/chemistry , DNA/genetics , DNA Restriction Enzymes , Molecular Sequence Data , Plasmids
9.
Biophys Chem ; 33(1): 55-61, 1989 Mar.
Article in English | MEDLINE | ID: mdl-2720090

ABSTRACT

Terbium (Tb3+) fluorescence was used to investigate local non-denaturation perturbations of double-helical DNA structure induced in this nucleic acid by various physical and chemical agents. It has been shown that the interaction of Tb3+ with DNA into which single-strand or double-strand breaks have been introduced by DNase I or by low doses of ionizing radiation does not influence the fluorescence of the lanthanide cation. On the other hand, interaction of terbium with DNA modified by the antitumour drug cis-diamminedichloroplatinum(II) at low levels of binding and by low doses of ultraviolet radiation (wavelength 254 nm) has been shown to result in substantial enhancement of the fluorescence of this cation. It has been proposed that the terbium fluorescent probe can also be exploited successfully for the purpose of analysing the guanine bases present in distorted double-stranded regions of DNA, in which only the vertical stacking of the base-pairs is altered.


Subject(s)
DNA , Nucleic Acid Conformation , Terbium , Antineoplastic Agents , Circular Dichroism , Cisplatin , DNA/radiation effects , Spectrometry, Fluorescence/methods , Ultraviolet Rays
10.
Gen Physiol Biophys ; 5(4): 423-32, 1986 Aug.
Article in English | MEDLINE | ID: mdl-3770461

ABSTRACT

Spectrophotometry, thermal denaturation, sedimentation, and viscometric techniques were used in a study of interaction of double helical DNA with an asymmetric phenazinium derivative, aposafranine. The results obtained indicate that aposafranine binds to DNA by a single binding mode, a wedge-like partial intercalation.


Subject(s)
DNA/drug effects , Intercalating Agents/pharmacology , Phenazines/pharmacology , Animals , Binding Sites/drug effects , Cattle , Nucleic Acid Conformation/drug effects , Spectrometry, Fluorescence/methods , Spectrum Analysis/methods , Viscosity
11.
Biochim Biophys Acta ; 867(1-2): 31-5, 1986 May 27.
Article in English | MEDLINE | ID: mdl-3707966

ABSTRACT

DNA fragments from chicken erythrocytes were modified by cis-diamminedichloroplatinum(II), its trans-isomer and chlorodiethylenetriaminoplatinum(II) chloride. The conformation of the modified DNA fragments in ethanolic solutions was studied by circular dichroism spectroscopy. Non-modified DNA adopted the A-form in 81% ethanol. The modification of DNA by the three platinum compounds inhibited the B to A transition of DNA induced by high concentrations of ethanol roughly to the same extent. The results support the view that the binding of the platinum complexes to B-DNA lowers the conformational freedom of DNA so that it cannot acquire the A-conformation.


Subject(s)
Cisplatin/analogs & derivatives , Cisplatin/metabolism , DNA/metabolism , Nucleic Acid Conformation , Animals , Chemical Phenomena , Chemistry , Chickens , Circular Dichroism , Erythrocytes/analysis , Ethanol , Solutions
12.
Gen Physiol Biophys ; 4(6): 597-608, 1985 Dec.
Article in English | MEDLINE | ID: mdl-4085785

ABSTRACT

The optical properties of the DNA complexes with divalent platinum compounds of the cis-diamine type differing both in the nature of anionic and neutral ligands and in the spatial arrangement about the platinum atom were studied. The platinum compounds cis-[Pt(NH3)2Cl2], [Pt(en)Cl2], [Pt(tetrameen)Cl2], cis-[Pt(NH3)2NO2Cl], and cis-[PtNH3(Bz)Cl2] at small values of r (r is the molar ratio of a platinum compound to DNA nucleotides in the reaction mixture) were found to induce an increase in the amplitude of the positive band in the circular dichroic (CD) spectrum of linear DNA. All the compounds listed except cis-[Pt(NH3)2NO2Cl] caused a sharp decrease of the amplitude of the negative band in the CD spectrum of a liquid crystalline microphase of DNA formed in solution in the presence of poly(ethylene glycol). All these platinum compounds (except [Pt(tetrameen)Cl2]) exhibit biological (antimitotic, antitumour, etc.) activity. The platinum compounds trans-[Pt(NH3)Cl2], trans-[Pt(NH3)2NO2Cl], cis-[PtNH3PyCl2], cis-[Pt(NH3)2(NO2)2], and [Pt(NH3)3Cl]Cl exhibiting a low (if any) biological activity, either induced a decrease of the amplitude of the positive band in the CD spectrum of linear DNA, or did not affect the CD spectrum at all. The effect of these platinum compounds on the CD spectrum of the liquid crystalline microphase of DNA was either weak or absent. It is assumed that the specific biological action of platinum compounds of the cis-diamine type is determined by the polydentate binding to DNA: in addition to the cis-bidentate covalent binding of platinum to DNA nitrogen bases, a hydrogen bond formation between the DNA and cis-amino ligands occurs by means of protons at nitrogen atoms.


Subject(s)
DNA , Nucleic Acid Conformation , Platinum , Circular Dichroism , Kinetics , Structure-Activity Relationship
13.
Antibiotiki ; 29(10): 743-8, 1984 Oct.
Article in Russian | MEDLINE | ID: mdl-6508249

ABSTRACT

The optical properties of the DNA complexes with the compounds of bivalent platinum were studied. The compounds differed by the nature of the anionic and neutral ligands and their spatial arrangement about the platinum atom. It was shown that the same as cis-[Pt (NH3)2Cl2] the platinum compounds with the biological activity, i.e. [Pt (en) Cl2], cis-[PtNH3 (Bz) Cl2] and cis-[Pt (NH3)2NO2Cl] induced at low values of r (a ratio of the number of the platinum moles added to the number of the DNA nucleotide moles in the solution) an increase in the amplitude of the positive band in the spectrum of the circular dichroism (CD) of the linear DNA and a marked decrease in the amplitude of the negative band in the spectrum of the CD of the liquid crystalline microphase of DNA formed in the presence of polyethyleneglycol. By the character of the action on the CD spectrum of the linear and condensed DNA [Pt (tetrameen)Cl2] which had no selective antimitotic effect might be referred to the above platinum compounds. Trans-[Pt (NH3)2NO2Cl], [PtNH3PyCl2], cis-[Pt (NH3)2(NO2)2] and [Pt (NH3)3Cl]Cl having no biological activity either induced only a decrease in the amplitude of the positive band in the CD spectrum of the linear DNA or had no effect on the CD spectrum. The effect of these compounds on the CD spectrum of the liquid crystalline microphase of DNA was slightly pronounced or not observed.


Subject(s)
Antineoplastic Agents/pharmacology , DNA/pharmacology , Organoplatinum Compounds/pharmacology , Animals , Antineoplastic Agents/analysis , Circular Dichroism , DNA/analysis , Ligands , Male , Molecular Weight , Organoplatinum Compounds/analysis , Salmon , Solutions , Spectrophotometry, Ultraviolet , Structure-Activity Relationship
15.
Folia Biol (Praha) ; 26(4): 275-88, 1980.
Article in English | MEDLINE | ID: mdl-7409270

ABSTRACT

Influence of fixation and so-called pretreatment procedures used in visualization of chromosome G-bands on staining capacity of isolated biopolymers in solution and on chromatin of cell nuclei was investigated. Whereas acid fixation as well as pretreatment procedures generally decreased the maximum binding capacity of isolated DNA and DNP for methylene blue, the dye content in acid-fixed nuclei was increased as compared with chromatin of neutral-fixed and non-fixed nuclei. The role of dehistonization and aggregation in an acid medium on the dye-binding properties of these two types of DNP was examined and the relation of higher-order structure of chromatin to its staining properties as reflected in G-banding pattern was discussed.


Subject(s)
Chromosome Banding/methods , DNA/metabolism , Deoxyribonucleoproteins/metabolism , Fixatives/pharmacology , Methylene Blue/metabolism , Nucleoproteins/metabolism , Acids , Animals , Cattle , Histocytochemistry
16.
Biophys Chem ; 9(2): 121-31, 1979 Jan.
Article in English | MEDLINE | ID: mdl-106900

ABSTRACT

Fluorescence and circular dichroism spectral measurements, thermal denaturation studies and binding competition experiments with netropsin and actinomycin D were carried out in systems containing phenosafranine bound to DNA's differing in base composition. The investigated properties exhibit a heterogeneity related to the content of A.T and G.C pairs in DNA and to the nature of phenosafranine binding modes. At low level of saturation of binding sites (r less than 0.1) phenosafranine does not show strong preference for any of the DNA base pairs in the overall binding. However, the strong monomer non-cooperative binding outside the helix (mode I1) occurs predominantly, even though not exclusively in G.C rich regions. The strong binding modes involving intercalated dye molecules (mode I2 and eventually mode II1) prevail in A.T rich regions. These binding modes become the principal types of strong phenosafranine interaction with DNA when the level of saturation of binding sites increases, i.e. at r greater than 0.1.20


Subject(s)
Coloring Agents , DNA , Phenazines , Bacillus cereus/analysis , Binding, Competitive , Chemical Phenomena , Chemistry , Circular Dichroism , DNA, Bacterial , Escherichia coli/analysis , Fluorescence , Hot Temperature , Micrococcus/analysis , Molecular Conformation , Netropsin , Nucleic Acid Denaturation , Osmolar Concentration
18.
Biophys Chem ; 8(1): 17-25, 1978 Mar.
Article in English | MEDLINE | ID: mdl-16996450

ABSTRACT

Aggregation or phenosafranine in concentrated aqueous solutions and its interaction with polyphosphates was Studied by absorption and fluorescence spectroscopy. At concentrations > 10(-3) M phenosafranine forms dimers (Kd = 3.8 x 10(2) l.mole(-1)), which are characterized by a hypsochromic shift of the visible and near ultraviolet absorption maxima accompanied by a hypochromic effect. No fluorescence could be detected from phenosafranine dimers. Analogous spectral changes were observed when a polyphosphate was titrated with phenusafranine, which indicated that with increasing saturation of the polyphosphate binding sites phenosafranine gradually became bound in the aggregated form. Full saturation of the polyphosphate binding sites with phenosafranine was reached only when an excess of free dye was present. The cooperative binding of phenosafranine to a polyphosphate could be evaluated by means of a theory proposed by Schwarz et al. At the zero ionic strength and at 25 degrees C the binding was characterized by cooperative binding constant K = 6.2 x 10(5) l.mole(-1), number of binding sites per monomeric phosphate residue g = 0.4, and cooperativity parameter q reverse similar 30. Spectroscopic properties of phenosafranine in the aggregated and poly phosphate-bound stotes were compared with those of ethidium bromide.

19.
Folia Biol (Praha) ; 12(3): 201-8, 1976.
Article in English | MEDLINE | ID: mdl-62683

ABSTRACT

Maximum binding capacity of calf thymus DNA and deoxyribonucleoprotein for methylene blue was investigated. The binding capacity of native and heat-denatured solutions of the biopolymers was compared with that of the solutions incubated with fixatives used in cytology, viz. 4% formaldehyde and methanol-acetic acid (3 : 1) mixture. The binding capacity of DNA and DNP is strongly reduced after the treatment with both fixation agents. Thermal denaturation leads to a small decrease in the binding capacity of DNA, but does not influence that of DNP. These results were compared with microphotometric determination of nuclear methylene blue content in thymocyte spreads. Fixation with MeAc increased the dye content as compared with air-dried thymocytes, whereas 4% formaldehyde exhibited practically no influence. The mechanisms connected with the different staining properties after incubating with fixation agents DNA and DNP solutions on the one hand and the nuclear material of thymocyte spreads on the other hand are discussed.


Subject(s)
DNA , Deoxyribonucleoproteins , Histological Techniques , Methylene Blue , Nucleoproteins , Binding Sites , Formaldehyde , Nucleic Acid Denaturation , Staining and Labeling , T-Lymphocytes
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