Unknown

Dataset Information

0

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 ?-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) repulsive interactions led to significantly increased (decreased) aggregation resistance, even based solely on single-point mutations. However, in the case of a particular amino acid (E17), the aggregation mechanism was altered by substitution with R or K, and this greatly mitigated improvements in aggregation resistance. The results illustrate that predictions based on native protein-protein interactions can provide a useful design target for engineering aggregation resistance; however, this approach needs to be balanced with consideration of how mutations can impact aggregation mechanisms.

SUBMITTER: O'Brien CJ 

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

REPOSITORIES: biostudies-literature

altmetric image

Publications

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

O'Brien C J CJ   Blanco M A MA   Costanzo J A JA   Enterline M M   Fernandez E J EJ   Robinson A S AS   Roberts C J CJ  

Protein engineering, design & selection : PEDS 20160509 6


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 effe  ...[more]

Similar Datasets

| S-EPMC4420528 | biostudies-literature
| S-EPMC6226484 | biostudies-literature
| S-EPMC3368139 | biostudies-literature
| S-EPMC4794390 | biostudies-literature
| S-EPMC2984139 | biostudies-literature
| S-EPMC2242593 | biostudies-literature
| S-EPMC7377432 | biostudies-literature
| S-EPMC4956399 | biostudies-literature
| S-EPMC6452746 | biostudies-literature
| S-EPMC6290019 | biostudies-literature