Effect of carbamylation on protein structure and adsorption to self-assembled monolayer surfaces

Abstract

Protein adsorption research has primarily focused upon the effects of surface chemistry, with almost no emphasis on how changes to proteins that occur in various disease states may influence their adsorption. One such situation occurs with chronic kidney disease where, despite hemodialysis treatment, the retention of urea within the blood compartment leads to protein carbamylation. Protein carbamylation has been shown to alter the function and structure of proteins. This work is focused on understanding how different degrees of carbamylation affect the physicochemical properties (structure, charge, water interactions) of single proteins (α-lactalbumin, albumin, and fibrinogen) and their adsorption to self-assembled monolayers. It was found that, unlike its secondary structure, the protein’s tertiary structure was significantly altered upon carbamylation. Also, compared to native proteins, an increase in carbamylation lead to an increase in the negative surface charge of the protein and a weaker hydration state of the protein. In order to study the effects of different types of neutral surfaces, of different surface-water properties, on protein adsorption both bare and alkanethiol modified (−CH3 or −OH end-groups) Au surfaces with were used as model surfaces. A significant decrease in adsorbed amounts of carbamylated fibrinogen and carbamylated α-lactalbumin, but not for carbamylated albumin, relative to native proteins was observed for both surfaces; suggesting that the increase in negative surface charge is more influential on adsorption than the change in hydration that occurs throughout the protein upon carbamylation. This data suggests that protein alterations that occur due to disease states have a significant effect on the overall protein structure and these changes affect their adsorption to surfaces.