PREPARATIVE CAPILLARY-ZONE-ELECTROPHORESIS IN COMBINATION WITH OFF-LINE GRAPHITE-FURNACE ATOMIC-ABSORPTION FOR THE ANALYSIS OF DNA COMPLEXES FORMED BY A NEW AMINOCOUMARINE PLATINUM (II) COMPOUND

Citation
Dld. Deforce et al., PREPARATIVE CAPILLARY-ZONE-ELECTROPHORESIS IN COMBINATION WITH OFF-LINE GRAPHITE-FURNACE ATOMIC-ABSORPTION FOR THE ANALYSIS OF DNA COMPLEXES FORMED BY A NEW AMINOCOUMARINE PLATINUM (II) COMPOUND, Electrophoresis, 19(14), 1998, pp. 2454-2458
Citations number
20
Categorie Soggetti
Biochemical Research Methods","Chemistry Analytical
Journal title
ISSN journal
01730835
Volume
19
Issue
14
Year of publication
1998
Pages
2454 - 2458
Database
ISI
SICI code
0173-0835(1998)19:14<2454:PCICWO>2.0.ZU;2-L
Abstract
Calf thymus DNA was incubated in vitro with a new aminocoumarin platin um(II) complex in order to study its interaction with DNA. The platina ted DNA was hydrolyzed enzymatically to the 5'-mononucleotide level us ing DNAase I and nuclease P1. Analysis of the DNA hydrolysate with cap illary zone electrophoresis (CZE), using sample stacking, revealed the presence of unhydrolyzed oligonucleotides in the platinated DNn. A ho memade system, using only some plastic pipet tips, was constructed to collect the oligonucleotide fraction during CZE analysis. The platinum content of this fraction was determined using graphite furnace atomic absorption with Zeeman background correction. This system proved to b e a useful tool to detect platinated DNA species (with a quantifiable detection limit for the detection of platinum of 0.78 ng). Subsequent gel filtration experiments confirmed the presence of high molecular we ight oligonucleotides that were platinated. This was proven by reversa l of the platination using thiourea and subsequent enzymatic hydrolysi s to 5'-mononucleotides.