A pH-responsive interface derived from resilin-mimetic protein Rec1- resilin

Abstract

In this investigation, for the first time we report the effects of pH on the molecular orientation, packing density, structural properties, adsorption characteristics and viscoelastic behaviour of resilin-mimetic protein rec1- resilin at the solid–liquid interface using quartz crystal microbalance with dissipation monitoring (QCM-D) and surface plasmon resonance (SPR) spectroscopy. QCM-D and SPR data confirm that the binding ability of rec1- resilin on a substrate is strongly pH-dependent the protein packing density on a gold surface is calculated to be 4.45 × 10 13 per cm 2 at the isoelectric point (IEP ∼ 4.9), 8.79 × 10 11 per cm 2 at pH 2 and 9.90 × 10 11 per cm 2 at pH 12, respectively. Our findings based on the thickness, dissipation and viscoelasticity of the rec1- resilin adlayer also indicate that it is adsorbed onto the gold substrate with different orientation depending on pH, such as back-on adsorption at acidic pH of 2, compact end-on bilayer adsorption at the IEP and side-on at high alkaline pH of 12. When rec1- resilin is ‘pinned’ to the substrate at IEP and subsequently exposed to an electrolyte solution adjusted to different pH, it switches from a compact globular conformation of the bio-macromolecule at the IEP to a coil conformation at pH between IEP to IED (IED = pKa value of tyrosine amino acid residue) and an extended coil conformation at pH > IED. This transformation from globule to coil to extended coil conformation is kinetically fast, robust and completely reversible. Such responsive surfaces created using ‘smart’ biomimetic rec1- resilin have the potential to find applications in many areas including biotechnology, medicine, sensors, controlled drug delivery systems and engineering.