Deep Mutational Scanning for the Study and Engineering of Protein Assemblies

Protein assemblies, such as fibers, cages, and sheets, are essential components of biological systems, with versatile functions that make them attractive engineering targets for biotechnological applications. Understanding the complex sequence–structure–function relationships that govern these assemblies is critical for both basic science and the engineering of novel nanomaterials. Deep mutational scanning (DMS) has emerged as a powerful technique for mapping these relationships across large sections of protein sequence space. Specifically, DMS couples high‐throughput assays with next‐generation sequencing technologies to create datasets that report on how changes to protein sequence alter protein function. This review provides an overview of protein assemblies and the basic principles of DMS, followed by a discussion of how DMS has been applied to protein assemblies, and what unique considerations arise when performing such studies. We aim to provide a comprehensive foundation for researchers across biochemistry and chemical biology looking to leverage such high‐throughput approaches to understand and engineer the next generation of protein‐based assemblies.