Knowledge-Based Unfolded State Model for Protein Design.

The design of proteins and miniproteins is an important challenge. Designed variants should be stable, meaning the folded/unfolded free energy difference should be large enough. Thus, the unfolded state plays a central role. An extended peptide model is often used, where side chains interact with solvent and nearby backbone, but not each other. The unfolded energy is then a function of sequence composition only and can be empirically parametrized. If the space of sequences is explored with a Monte Carlo procedure, protein variants will be sampled according to a well-defined Boltzmann probability distribution. We can then choose unfolded model parameters to maximize the probability of sampling native-like sequences. This leads to a well-defined maximum likelihood framework. We present an iterative algorithm that follows the likelihood gradient. The method is presented in the context of our Proteus software, as a detailed downloadable tutorial. The unfolded model is combined with a folded model that uses molecular mechanics and a Generalized Born solvent. It was optimized for three PDZ domains and then used to redesign them. The sequences sampled are native-like and similar to a recent PDZ design study that was experimentally validated.