Unique supramolecular assembly of a redox protein with nucleic acids onto hybrid bilayer: towards a dynamic DNA chip.
Highly controlled supramolecular assemblies combining a genetically engineered redox protein, cytochrome b5, and modified oligonucleotides are presented. Modified b5 and DNA are covalently assembled through a hetero bifunctional cross-linker to give a unique hybrid molecular species. Moreover, the assembly includes a histidine tag head able to bind to modified phospholipids which lead to a new generation of self-assembled dynamic DNA chips. The interaction of the construction with a complementary oligonucleotide sequence can be monitored in real time by surface plasmon resonance using Biacore technology. The biochip, presented herein, features unique properties including tunable surface density of probes, very low non-specific interactions and optimization of hybridization efficiency. In addition, we demonstrated that the phase transition of the lipidic layer can modulate the dynamic of the association of the complex to the supported membrane. Potential applications of this new device are multiple including high sensitivity and high selectivity biochips, especially for studies of the DNA-ligands interactions in a biomimetic environment.
Boireau
W
W
Institut FEMTO-ST, Département LPMO, CNRS, UMR 6174, F-25044 Besançon Cedex, France. wboireau@lpmo.edu
Zeeh
J C
JC
Puig
P E
PE
Pompon
D
D
D015786:Cytochromes b5 / Q000032:analysis* / Q000737:chemistry*; D004247:DNA / Q000032:analysis* / Q000737:chemistry*; D004268:DNA-Binding Proteins / Q000032:analysis / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry*; D046911:Macromolecular Substances / Q000032:analysis / Q000737:chemistry; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010084:Oxidation-Reduction; D010743:Phospholipids / Q000737:chemistry; D011485:Protein Binding; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000379:methods*
D004268:DNA-Binding Proteins; D008051:Lipid Bilayers; D046911:Macromolecular Substances; D010743:Phospholipids; D004247:DNA; D015786:Cytochromes b5