Analysis of mismatched DNA by mismatch binding ligand (MBL)-Sepharose affinity chromatography.
Mismatch binding molecules (MBLs), strongly and selectively bound to the mismatched base pair in duplex DNA, were immobilized on Sepharose. Three MBL-Sepharose columns were prepared with three MBLs, naphthyridine dimer (ND), naphthyridine-azaquinolone (NA), and aminonaphthyridine dimer (amND), which exhibited different binding profiles to the mismatched base pairs. These three MBL-Sepharose columns showed characteristic elution profiles for DNA duplexes containing mismatched base pairs. The ND-Sepharose column separated the G-G and G-A mismatched DNA from fully matched duplexes. The NA-Sepharose column separated the A-A and G-A mismatched DNA from other DNA duplexes. The amND-Sepharose column separated the C-C mismatched DNA. These chromatographic profiles were very consistent with the binding preference of each MBL. By changing the elution conditions from sodium hydroxide to sodium chloride, MBL-Sepharose columns were also able to separate the mismatched DNA that weakly bound to the MBL from fully matched DNA duplex. Figure MBL-Sepharose affinity chromatography successfully separates the mismatched duplex DNA from fully matched duplex.
Goto
Yuki
Y
Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto, Japan.
Suda
Hitoshi
H
Kobori
Akio
A
Nakatani
Kazuhiko
K
D020137:Base Pair Mismatch; D002196:Capillaries; D002846:Chromatography, Affinity / Q000379:methods*; D002847:Chromatography, Agarose / Q000379:methods*; D004247:DNA / Q000737:chemistry*; D019281:Dimerization; D008024:Ligands; D008956:Models, Chemical; D009154:Mutation; D009287:Naphthyridines / Q000032:analysis; D020641:Polymorphism, Single Nucleotide; D015363:Quinolones / Q000032:analysis; D012685:Sepharose / Q000737:chemistry; D012965:Sodium Chloride / Q000737:chemistry; D013696:Temperature
D008024:Ligands; D009287:Naphthyridines; D015363:Quinolones; D012965:Sodium Chloride; D004247:DNA; D012685:Sepharose