title: Molecular recognition of oligonucleotides and plasmid DNA by l-methionine.
abstract: The present study explores the effect of oligonucleotide composition on the mechanism of retention to l-methionine agarose support by chromatography and saturation transfer difference (STD)-nuclear magnetic resonance (NMR) techniques. All chromatographic experiments were performed using 1.5 M (NH4 )2 SO4 . The binding profiles obtained by chromatography show that oligonucleotides with thymine had the highest retention time. In general, the larger homo-oligonucleotides are more retained to the l-methionine agarose support. Moreover, the study with hetero-oligonucleotides confirms that the presence of guanine reduces the retention on the l-methionine chromatographic support. These results are in accord with STD-NMR experiments, which show that the strongest signals were observed for the methyl group of thymine, and no STD signals were observed for the guanosine protons. Finally, the retention behaviour of linear plasmid DNA (pDNA) with different sizes and base composition (2.7-kbp pUC19, 6.05-kbp pVAX1-LacZ, 7.4-kbp pVAX1-LacZgag and 14-kbp pcDNA-based plasmid) was also evaluated by chromatography. The results indicate that the underlying mechanism of retention involves not only hydrophobic interactions but also other elementary interactions responsible for the biorecognition of pDNA molecules by l-methionine ligands.
authors.affiliation: CICS-UBI-Centro de Investigação em Ciências da Saúde, Universidade da Beira Interior, Av. Infante D. Henrique, 6200-506, Covilhã, Portugal.
authors.forename: Elia
authors.initials: E
authors.lastname: Mota
authors.forename: Fani
authors.initials: F
authors.lastname: Sousa
authors.forename: Angela
authors.initials: A
authors.lastname: Sousa
authors.forename: João A
authors.initials: JA
authors.lastname: Queiroz
authors.forename: Carla
authors.initials: C
authors.lastname: Cruz
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D004247:DNA / Q000737:chemistry*; D008715:Methionine / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009841:Oligonucleotides / Q000737:chemistry*; D010957:Plasmids / Q000235:genetics
chemical_list: D009841:Oligonucleotides; D004247:DNA; D008715:Methionine
keywords: l-methionine agarose support; molecular recognition; oligonucleotides; plasmid DNA isoforms; saturation transfer difference nuclear magnetic resonance