Highly sensitive electrochemiluminescence displacement method for the study of DNA/small molecule binding interactions
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| Title: | Highly sensitive electrochemiluminescence displacement method for the study of DNA/small molecule binding interactions |
|---|---|
| Authors: | Huang, Rongfu1, Wang, Li-Rong1, Guo, Liang-Hong LHGuo@rcees.ac.cn |
| Source: | Analytica Chimica Acta. Aug2010, Vol. 676 Issue 1/2, p41-45. 5p. |
| Subjects: | Chemiluminescence, DNA, Genetic regulation, Molecular self-assembly, Binding sites, Ligand binding (Biochemistry), Molecules |
| Abstract: | Abstract: Non-covalent binding interactions of small molecules with DNA play important roles in regulating gene expression and gene function. In this work, a highly sensitive electrochemiluminescence (ECL) displacement method has been developed to investigate such interactions, particularly for weak DNA binders. This ECL method relies on a double-stranded DNA film deposited on an indium tin oxide electrode (ITO) surface by layer-by-layer self-assembly. A DNA intercalator, [Ru(bpy)2(dppz)]2+ (bpy=2,2′-bipyridine, dppz=dipyrido[3,2-a:2′3′-c]phenazine), is employed as the ECL signal indicator. If a test compound competes with the indicator for the same binding sites in DNA, it would displace the indicator from the film and reduce ECL signal. The new method was validated by measuring five well-known DNA-binding organic molecules including quinacrine, H33258, thiazole orange, ethidium bromide and 4,6-diamidine-2-phenylindole dihydrochloride. Due to high ECL sensitivity, only 0.4μmolL−1 [Ru(bpy)2(dppz)]2+ was needed in the ECL displacement measurement, which is about 75-fold less than the concentration in the voltammetric measurement. The lowered concentration permitted direct measurement of IC50 values of eight hydroxylated polycyclic aromatic hydrocarbons in their ECL displacement curves and subsequent calculation of their binding constants with DNA. The ECL displacement method is particularly useful for investigating weak DNA binders with limited aqueous solubility. [ABSTRACT FROM AUTHOR] |
| Copyright of Analytica Chimica Acta is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) | |
| Database: | Engineering Source |
| FullText | Text: Availability: 0 |
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| Header | DbId: egs DbLabel: Engineering Source An: 53335423 AccessLevel: 6 PubType: Academic Journal PubTypeId: academicJournal PreciseRelevancyScore: 0 |
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| Items | – Name: Title Label: Title Group: Ti Data: Highly sensitive electrochemiluminescence displacement method for the study of DNA/small molecule binding interactions – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Huang%2C+Rongfu%22">Huang, Rongfu</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Wang%2C+Li-Rong%22">Wang, Li-Rong</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Guo%2C+Liang-Hong%22">Guo, Liang-Hong</searchLink><i> LHGuo@rcees.ac.cn</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22Analytica+Chimica+Acta%22">Analytica Chimica Acta</searchLink>. Aug2010, Vol. 676 Issue 1/2, p41-45. 5p. – Name: Subject Label: Subjects Group: Su Data: <searchLink fieldCode="DE" term="%22Chemiluminescence%22">Chemiluminescence</searchLink><br /><searchLink fieldCode="DE" term="%22DNA%22">DNA</searchLink><br /><searchLink fieldCode="DE" term="%22Genetic+regulation%22">Genetic regulation</searchLink><br /><searchLink fieldCode="DE" term="%22Molecular+self-assembly%22">Molecular self-assembly</searchLink><br /><searchLink fieldCode="DE" term="%22Binding+sites%22">Binding sites</searchLink><br /><searchLink fieldCode="DE" term="%22Ligand+binding+%28Biochemistry%29%22">Ligand binding (Biochemistry)</searchLink><br /><searchLink fieldCode="DE" term="%22Molecules%22">Molecules</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: Abstract: Non-covalent binding interactions of small molecules with DNA play important roles in regulating gene expression and gene function. In this work, a highly sensitive electrochemiluminescence (ECL) displacement method has been developed to investigate such interactions, particularly for weak DNA binders. This ECL method relies on a double-stranded DNA film deposited on an indium tin oxide electrode (ITO) surface by layer-by-layer self-assembly. A DNA intercalator, [Ru(bpy)2(dppz)]2+ (bpy=2,2′-bipyridine, dppz=dipyrido[3,2-a:2′3′-c]phenazine), is employed as the ECL signal indicator. If a test compound competes with the indicator for the same binding sites in DNA, it would displace the indicator from the film and reduce ECL signal. The new method was validated by measuring five well-known DNA-binding organic molecules including quinacrine, H33258, thiazole orange, ethidium bromide and 4,6-diamidine-2-phenylindole dihydrochloride. Due to high ECL sensitivity, only 0.4μmolL−1 [Ru(bpy)2(dppz)]2+ was needed in the ECL displacement measurement, which is about 75-fold less than the concentration in the voltammetric measurement. The lowered concentration permitted direct measurement of IC50 values of eight hydroxylated polycyclic aromatic hydrocarbons in their ECL displacement curves and subsequent calculation of their binding constants with DNA. The ECL displacement method is particularly useful for investigating weak DNA binders with limited aqueous solubility. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of Analytica Chimica Acta is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
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| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1016/j.aca.2010.07.033 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 5 StartPage: 41 Subjects: – SubjectFull: Chemiluminescence Type: general – SubjectFull: DNA Type: general – SubjectFull: Genetic regulation Type: general – SubjectFull: Molecular self-assembly Type: general – SubjectFull: Binding sites Type: general – SubjectFull: Ligand binding (Biochemistry) Type: general – SubjectFull: Molecules Type: general Titles: – TitleFull: Highly sensitive electrochemiluminescence displacement method for the study of DNA/small molecule binding interactions Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Huang, Rongfu – PersonEntity: Name: NameFull: Wang, Li-Rong – PersonEntity: Name: NameFull: Guo, Liang-Hong IsPartOfRelationships: – BibEntity: Dates: – D: 31 M: 08 Text: Aug2010 Type: published Y: 2010 Identifiers: – Type: issn-print Value: 00032670 Numbering: – Type: volume Value: 676 – Type: issue Value: 1/2 Titles: – TitleFull: Analytica Chimica Acta Type: main |
| ResultId | 1 |
