Zero-shot protein stability prediction by inverse folding models: a free energy interpretation

Jes Frellsen (Technical University of Denmark) · Maher Kassem (University of Copenhagen) · Tone Bengtsen (ADC Consulting) · Lars Olsen (Novonesis) · Kresten Lindorff-Larsen (University of Copenhagen) · Jesper Ferkinghoff-Borg (Novo Nordisk A/S) · Wouter Boomsma (University of Copenhagen)
amino acid preferencesapproximation methodsempirical assessmentfree-energy considerationsinverse folding modelslikelihood ratiosperformance gainspredictive modelingprotein stabilityrelative stabilitystability estimationstability predictiontheoretical perspectivethermodynamic stabilityzero-shot predictors

Inverse folding models have proven to be highly effective zero-shot predictors of protein stability. Despite this success, the link between the amino acid preferences of an inverse folding model and the free-energy considerations underlying thermodynamic stability remains incompletely understood. A better understanding would be of interest not only from a theoretical perspective, but also potentially provide the basis for stronger zero-shot stability prediction. In this paper, we take steps to clarify the free-energy foundations of inverse folding models. Our derivation reveals the standard practice of likelihood ratios as a simplistic approximation and suggests several paths towards better estimates of the relative stability. We empirically assess these approaches and demonstrate that considerable gains in zero-shot performance can be achieved with fairly simple means.