Weak self-interactions of globular proteins studied by small-angle X-ray scattering and structure-based modeling

TL;DR

This study combines SAXS experiments and theoretical modeling to analyze weak protein-protein interactions in solution, revealing that hard-core and electrostatic forces primarily determine the structure factor, with other soft interactions being negligible or canceling out.

Contribution

It provides a comprehensive analysis of protein interactions using combined experimental and modeling approaches, highlighting the dominant role of hard-core and electrostatic interactions and the effects of charge asymmetry.

Findings

Structure factor mainly governed by hard-core and electrostatic interactions.

Soft interactions like van der Waals are insignificant or cancel out.

Charge asymmetry influences electrostatic repulsion and shape effects.

Abstract

We investigate protein-protein interactions in solution by small-angle X-ray scattering (SAXS) and theoretical modeling. The structure factor for solutions of bovine pancreatic trypsin inhibitor (BPTI), myoglobin (Mb), and intestinal fatty acid-binding protein (IFABP) is determined from SAXS measurements at multiple concentrations, from Monte Carlo simulations with a coarse-grained structure-based interaction model, and from analytic approximate solutions of two idealized colloidal interaction models without adjustable parameters. By combining these approaches, we find that the structure factor is essentially determined by hard-core and screened electrostatic interactions. Other soft short-ranged interactions (van der Waals and solvation-related) are either individually insignificant or tend to cancel out. The structure factor is also not significantly affected by charge fluctuations. For Mb and IFABP, with small net charge and relatively symmetric charge distribution, the structure factor is well described by a hard-sphere model. For BPTI, with larger net charge, screened electrostatic repulsion is also important, but the asymmetry of the charge distribution reduces the repulsion from that predicted by a charged hard-sphere model with the same net charge. Such charge asymmetry may also amplify the effect of shape asymmetry on the protein-protein potential of mean force.