SURFACE-PLASMON RESONANCE BASED KINETIC-STUDIES OF ZINC-FINGER DNA INTERACTIONS

Libraries of the zinc finger DNA binding protein, Zif268, have been co nstructed and selected for affinity and specificity toward DNA targets using the phage display technique (Wu et al., 1995). Mutant proteins were purified to homogeneity and were characterized for their ability to interact with their DNA targets using a real-time biomolecular inte raction assay (BIA). One mutant protein, C7, bound the Zif268 consensu s binding sequence with a 13-fold increase in affinity as compared to the wild-type Zif268 protein. Mutant proteins with moderate affinity f or new DNA targets within a consensus sequence of HIV-1 have also been obtained. Surface plasmon resonance based BLA has provided invaluable kinetic information which offers insights into the mechanism of prote in-DNA interactions.