Tuning protein GlnB-Hs surface interaction with silicon: FTIR-ATR, AFM and XPS study

[Display omitted] ► Thin films of GlnB-Hs protein were prepared on hydrophilic and hydrophobic silicon. ► GlnB-Hs is adsorbed face-up on hydrophilic silicon deprotonating its residues. ► On hydrophobic silicon, GlnB-Hs forms filaments adopting side-on conformation. ► GlnB-Hs secondary structure was conserved after adsorption on both surfaces. Herbaspirillum seropedicae GlnB (GlnB-Hs) is a signal transduction protein involved in the control of nitrogen, carbon and energetic metabolism. The adsorption of GlnB-Hs deposited by spin coating on hydrophilic and hydrophobic silicon forms a thin layer that was characterized using atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared attenuated total reflectance spectroscopy (FTIR-ATR). AFM allowed the identification of globular, face-up donut like array of protein on hydrophilic silicon substrate, favoring deprotonated residues to contact the silicon oxide surface. Over hydrophobic silicon, GlnB-Hs adopts a side-on conformation forming a filament network, avoiding the contact of protonated residues with silicon surface. XPS allowed us to determine the protonated and non-protonated states of nitrogen 1s (N 1s). The FTIR-ATR measurements provided information about protein secondary structure and its conservation, after surface adsorption.