Estimation of Binding Constants of Peptide Nucleic Acid and Secondary-Structured DNA by Affinity Capillary Electrophoresis
| Autor: | Tohru Takarada, Harumi Tsukada, Lal Mohan Kundu, Mizuo Maeda, Yukiharu Matsuoka, Naoki Kanayama |
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| Rok vydání: | 2012 |
| Předmět: |
Peptide Nucleic Acids
Polyethylene Glycols Analytical Chemistry chemistry.chemical_compound Nucleic acid thermodynamics Capillary electrophoresis Nucleotide Strand invasion chemistry.chemical_classification Base Sequence Peptide nucleic acid musculoskeletal neural and ocular physiology Inverted Repeat Sequences Electrophoresis Capillary Nucleic Acid Hybridization Affinity Labels DNA Binding constant Electrophoresis chemistry Biochemistry biological sciences cardiovascular system Biophysics Nucleic Acid Conformation Thermodynamics tissues |
| Zdroj: | Analytical Chemistry. 84:5204-5209 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/ac301025m |
| Popis: | An affinity capillary electrophoresis method was developed to determine a binding constant between a peptide nucleic acid (PNA) and a hairpin-structured DNA. A diblock copolymer composed of PNA and polyethylene glycol (PEG) was synthesized as a novel affinity probe. The base sequence of the probe's PNA segment was complementary to a hairpin-structured region of a 60-base single-stranded DNA (ssDNA). Upon applying a voltage, the DNA hairpin migrated slowly compared to a random sequence ssDNA in the presence of the PNA probe. This retardation was induced by strand invasion of the PNA into the DNA hairpin to form a hybridized complex, where the PEG segment received a large amount of hydrodynamic friction during electrophoresis. The binding constant between the PNA probe and the DNA hairpin was easily determined by mobility analysis. This simple method would be potentially beneficial in studying binding behaviors of various artificial nucleotides to natural DNA or RNA. |
| Databáze: | OpenAIRE |
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