Smart polymer-streptavidin conjugates.

Abstract

The conjugation of stimuli-responsive, or "smart," polymers to streptavidin is described. The polymer is synthesized with a thiol-reactive end-group, which is used to end-graft the polymer to cysteine or lysine side-chains that are genetically engineered into controlled positions on the streptavidin surface. The conjugation positions are chosen on the basis of their location relative to the binding site, together with the criteria that they be solvent accessible and thus reactive. The polymer composition can be controlled to impart responsiveness to temperature, pH, and/or specific wavelengths of light. These signals are sent to the polymer, which serves as an antennae and actuator to gate biotin or biotinylated protein association with the streptavidin binding sites. The molecular switching and gating activity is directed by the reversible polymer transition between a hydrophilic, expanded coil and a more hydrophobic, collapsed state that is smaller in volume. The differences in the polymer steric properties serve to block or allow ligand access to the binding site. The control of polymer molecular weight is a particularly important design parameter for these molecular gates.