ACMA (9-amino-6-chloro-2-methoxy acridine) forms three complexes in the presence of DNA

Natalia Busto, Begoña García, José M. Leal, Jorge F. Gaspar, Célia Martins, Alessia Boggioni, Fernando Secco

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The interaction of ACMA (9-amino-6-chloro-2-methoxy acridine) (D) with DNA (P) has been studied by absorbance, fluorescence, circular dichroism, spectrophotometry, viscometry and unwinding electrophoresis. A T-jump kinetic study has also been undertaken. The experimental data show that, totally unlike other drugs, ACMA is able to form with DNA three complexes (PD I, PD II, PD III) that differ from each other by the characteristics and extent of the binding process. The main features of PD I fulfil the classical intercalation pattern and the formation/dissociation kinetics have been elucidated by T-jump techniques. PD II and PD III are also intercalated species but, in addition to the dye units lodged between base pairs, they also bear dye molecules externally bound, more in PD III relative to PD II. A reaction mechanism is put forward here. Comparison between absorbance, fluorescence and kinetic experiments has enabled us to determine the binding constants of the three complexes, namely (6.5 ± 1.1) × 10 4 M -1 (PD I), (5.5 ± 1.5) × 10 4 M -1 (PD II) and (5.7 ± 0.03) × 10 4 M -1 (PD III). The Comet assay reveals that the ACMA binding to DNA brings about genotoxic properties. The mutagenic potential studied by the Ames test reveals that ACMA can produce frameshift and transversion/transition mutations. ACMA also is able to produce base-pair substitution in the presence of S9 mix. Moreover, the MTT assays have revealed cytotoxicity. The biological effects observed have been rationalized in light of these features.

Original languageEnglish
Pages (from-to)19534-19545
Number of pages12
JournalPhysical Chemistry Chemical Physics
Volume13
Issue number43
DOIs
Publication statusPublished - 21 Nov 2011

Fingerprint

Acridines
deoxyribonucleic acid
DNA
kinetics
dyes
viscometry
fluorescence
biological effects
Kinetics
spectrophotometry
electrophoresis
mutations
Assays
bears
comets
intercalation
Coloring Agents
dichroism
Fluorescence
drugs

Cite this

Busto, Natalia ; García, Begoña ; Leal, José M. ; Gaspar, Jorge F. ; Martins, Célia ; Boggioni, Alessia ; Secco, Fernando. / ACMA (9-amino-6-chloro-2-methoxy acridine) forms three complexes in the presence of DNA. In: Physical Chemistry Chemical Physics. 2011 ; Vol. 13, No. 43. pp. 19534-19545.
@article{8353dafe79624e73b454ee81f86c374f,
title = "ACMA (9-amino-6-chloro-2-methoxy acridine) forms three complexes in the presence of DNA",
abstract = "The interaction of ACMA (9-amino-6-chloro-2-methoxy acridine) (D) with DNA (P) has been studied by absorbance, fluorescence, circular dichroism, spectrophotometry, viscometry and unwinding electrophoresis. A T-jump kinetic study has also been undertaken. The experimental data show that, totally unlike other drugs, ACMA is able to form with DNA three complexes (PD I, PD II, PD III) that differ from each other by the characteristics and extent of the binding process. The main features of PD I fulfil the classical intercalation pattern and the formation/dissociation kinetics have been elucidated by T-jump techniques. PD II and PD III are also intercalated species but, in addition to the dye units lodged between base pairs, they also bear dye molecules externally bound, more in PD III relative to PD II. A reaction mechanism is put forward here. Comparison between absorbance, fluorescence and kinetic experiments has enabled us to determine the binding constants of the three complexes, namely (6.5 ± 1.1) × 10 4 M -1 (PD I), (5.5 ± 1.5) × 10 4 M -1 (PD II) and (5.7 ± 0.03) × 10 4 M -1 (PD III). The Comet assay reveals that the ACMA binding to DNA brings about genotoxic properties. The mutagenic potential studied by the Ames test reveals that ACMA can produce frameshift and transversion/transition mutations. ACMA also is able to produce base-pair substitution in the presence of S9 mix. Moreover, the MTT assays have revealed cytotoxicity. The biological effects observed have been rationalized in light of these features.",
author = "Natalia Busto and Bego{\~n}a Garc{\'i}a and Leal, {Jos{\'e} M.} and Gaspar, {Jorge F.} and C{\'e}lia Martins and Alessia Boggioni and Fernando Secco",
year = "2011",
month = "11",
day = "21",
doi = "10.1039/c1cp22158b",
language = "English",
volume = "13",
pages = "19534--19545",
journal = "Physical Chemistry Chemical Physics",
issn = "1463-9076",
publisher = "ROYAL SOC CHEMISTRY",
number = "43",

}

ACMA (9-amino-6-chloro-2-methoxy acridine) forms three complexes in the presence of DNA. / Busto, Natalia; García, Begoña; Leal, José M.; Gaspar, Jorge F.; Martins, Célia; Boggioni, Alessia; Secco, Fernando.

In: Physical Chemistry Chemical Physics, Vol. 13, No. 43, 21.11.2011, p. 19534-19545.

Research output: Contribution to journalArticle

TY - JOUR

T1 - ACMA (9-amino-6-chloro-2-methoxy acridine) forms three complexes in the presence of DNA

AU - Busto, Natalia

AU - García, Begoña

AU - Leal, José M.

AU - Gaspar, Jorge F.

AU - Martins, Célia

AU - Boggioni, Alessia

AU - Secco, Fernando

PY - 2011/11/21

Y1 - 2011/11/21

N2 - The interaction of ACMA (9-amino-6-chloro-2-methoxy acridine) (D) with DNA (P) has been studied by absorbance, fluorescence, circular dichroism, spectrophotometry, viscometry and unwinding electrophoresis. A T-jump kinetic study has also been undertaken. The experimental data show that, totally unlike other drugs, ACMA is able to form with DNA three complexes (PD I, PD II, PD III) that differ from each other by the characteristics and extent of the binding process. The main features of PD I fulfil the classical intercalation pattern and the formation/dissociation kinetics have been elucidated by T-jump techniques. PD II and PD III are also intercalated species but, in addition to the dye units lodged between base pairs, they also bear dye molecules externally bound, more in PD III relative to PD II. A reaction mechanism is put forward here. Comparison between absorbance, fluorescence and kinetic experiments has enabled us to determine the binding constants of the three complexes, namely (6.5 ± 1.1) × 10 4 M -1 (PD I), (5.5 ± 1.5) × 10 4 M -1 (PD II) and (5.7 ± 0.03) × 10 4 M -1 (PD III). The Comet assay reveals that the ACMA binding to DNA brings about genotoxic properties. The mutagenic potential studied by the Ames test reveals that ACMA can produce frameshift and transversion/transition mutations. ACMA also is able to produce base-pair substitution in the presence of S9 mix. Moreover, the MTT assays have revealed cytotoxicity. The biological effects observed have been rationalized in light of these features.

AB - The interaction of ACMA (9-amino-6-chloro-2-methoxy acridine) (D) with DNA (P) has been studied by absorbance, fluorescence, circular dichroism, spectrophotometry, viscometry and unwinding electrophoresis. A T-jump kinetic study has also been undertaken. The experimental data show that, totally unlike other drugs, ACMA is able to form with DNA three complexes (PD I, PD II, PD III) that differ from each other by the characteristics and extent of the binding process. The main features of PD I fulfil the classical intercalation pattern and the formation/dissociation kinetics have been elucidated by T-jump techniques. PD II and PD III are also intercalated species but, in addition to the dye units lodged between base pairs, they also bear dye molecules externally bound, more in PD III relative to PD II. A reaction mechanism is put forward here. Comparison between absorbance, fluorescence and kinetic experiments has enabled us to determine the binding constants of the three complexes, namely (6.5 ± 1.1) × 10 4 M -1 (PD I), (5.5 ± 1.5) × 10 4 M -1 (PD II) and (5.7 ± 0.03) × 10 4 M -1 (PD III). The Comet assay reveals that the ACMA binding to DNA brings about genotoxic properties. The mutagenic potential studied by the Ames test reveals that ACMA can produce frameshift and transversion/transition mutations. ACMA also is able to produce base-pair substitution in the presence of S9 mix. Moreover, the MTT assays have revealed cytotoxicity. The biological effects observed have been rationalized in light of these features.

UR - http://www.scopus.com/inward/record.url?scp=80055029272&partnerID=8YFLogxK

U2 - 10.1039/c1cp22158b

DO - 10.1039/c1cp22158b

M3 - Article

VL - 13

SP - 19534

EP - 19545

JO - Physical Chemistry Chemical Physics

JF - Physical Chemistry Chemical Physics

SN - 1463-9076

IS - 43

ER -