Interaction of Human Serum Albumin with short Polyelectrolytes: A study by Calorimetry and Computer Simulation

TL;DR

This study combines calorimetry and computer simulations to investigate how human serum albumin interacts with short poly(acrylic acid) chains, revealing thermodynamic details and binding mechanisms in aqueous solutions.

Contribution

It provides a detailed thermodynamic and mechanistic analysis of HSA-polymer interactions, validated by simulations that elucidate the binding site and driving forces.

Findings

Binding of one PAA chain per HSA molecule.

Binding free energy increases with temperature.

Counterion release drives the binding process.

Abstract

We present a comprehensive study of the interaction of human serum albumin (HSA) with poly(acrylic acid) (PAA; number average degree of polymerization: 25) in aqueous solution. The interaction of HSA with PAA is studied in dilute solution as the function of the concentration of added salt (20 - 100 mM) and temperature (25 - 37$^{\circ}$C). Isothermal titration calorimetry (ITC) is used to analyze the interaction and to determine the binding constant and related thermodynamic data. It is found that only one PAA chain is bound per HSA molecule. The free energy of binding $\Delta G_b$ increases with temperature significantly. $\Delta G_b$ decreases with increasing salt concentration and is dominated by entropic contributions due to the release of bound counterions. Coarse-grained Langevin computer simulations treating the counterions in an explicit manner are used study the process of binding in detail. These simulations demonstrate that the PAA chains are bound in the Sudlow II site of the HSA. Moreover, $\Delta G_b$ is calculated from the simulations and found to be in very good agreement with the measured data. The simulations demonstrate clearly that the driving force of binding is the release of counterions in full agreement with the ITC-data.