Modulating non-native aggregation and electrostatic protein-protein interactions with computationally designed single-point mutations.

ABSTRACT: Non-native protein aggregation is a ubiquitous challenge in the production, storage and administration of protein-based biotherapeutics. This study focuses on altering electrostatic protein-protein interactions as a strategy to modulate aggregation propensity in terms of temperature-dependent aggregation rates, using single-charge variants of human γ-D crystallin. Molecular models were combined to predict amino acid substitutions that would modulate protein-protein interactions with minimal effects on conformational stability. Experimental protein-protein interactions were quantified by the Kirkwood-Buff integrals (G22) from laser scattering, and G22 showed semi-quantitative agreement with model predictions. Experimental initial-rates for aggregation showed that increased (decreased) repul

SUBMITTER: O'Brien CJ 

PROVIDER: S-EPMC4867096 | biostudies-literature | 2016 Jun

REPOSITORIES: biostudies-literature