Binding mechanism of oligopeptides on solid surface: assessing the significance of single-molecule approach

This paper addresses the complementarity and potential disparities between single-molecule and ensemble-average approaches to probe the binding mechanism of oligopeptides on inorganic solids. Specifically, we explore the peptide/gold interface owing to its significance in various topics and its suitability to perform experiments both in model and real conditions. Experimental results show that the studied peptide adopts a lying configuration upon adsorption on the gold surface and interacts through its peptidic links and deprotonated thiolate extremities, in agreement with theoretical predictions. Single-molecule force spectroscopy (SMFS) measurements revealed the existence of a wide panel of adhesion forces, resulting from the interaction between individual peptide moieties and the abundant surface sites. We therefore propose methodological developments for sorting the events of interest to understand the peptide adsorption mechanism. Thermodynamic and kinetic aspects of the peptide adsorption are probed using both static and dynamic force spectroscopy measurements. Specifically, we show the possibility of providing a reasonable estimate of the peptide free energy of adsorption Δ_adsG° by exploring the fluctuations of the adhesion work, based on the Jarzynski equality, and by using a parametric Gamma estimator. The proposed approach offers a relevant method for studying the different factors influencing the peptide adsorption and evaluating their impact on Δ_adsG° as an alternative to exploring adhesion forces that may lead to misinterpretations. This is illustrated by the comparison of the adsorption of two peptides with specific amino acids substitution. Our method provides insights into the overall mechanism by which peptides interact with the surface and allows an integration of the single-molecule versus ensemble-average points of view.