Protein structure is tightly intertwined with function according to the laws of evolution. Understanding how structure determines function has been the aim of structural biology for decades. Here, we have wondered instead whether it is possible to exploit the function for which a protein was evolutionary selected to gain information on protein structure and on the landscape explored during the early stages of molecular and natural evolution. To answer to this question, we developed a new methodology, which we named CAMELS (Coupling Analysis by Molecular Evolution Library Sequencing), that is able to obtain the in vitro evolution of a protein from an artificial selection based on function. We were able to observe with CAMELS many features of the TEM-1 beta-lactamase local fold exclusively by generating and sequencing large libraries of mutational variants. We demonstrated that we can, whenever a functional phenotypic selection of a protein is available, sketch the structural and evolutionary landscape of a protein without utilizing purified proteins, collecting physical measurements, or relying on the pool of natural protein variants.

中文翻译：

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体外进化的蛋白质结构信息和进化景观。

根据进化规律，蛋白质结构与功能紧密相连。几十年来，了解结构如何决定功能一直是结构生物学的目标。在这里，我们想知道是否有可能利用蛋白质进化选择的功能来获取有关蛋白质结构以及分子和自然进化早期阶段探索的景观的信息。为了回答这个问题，我们开发了一种新的方法，我们将其命名为CAMELS（分子进化库测序耦合分析），该方法能够通过基于功能的人工选择来获得蛋白质的体外进化。通过生成大型突变体文库并对其进行测序，我们能够使用 CAMELS 观察 TEM-1 β-内酰胺酶局部折叠的许多特征。我们证明，每当蛋白质的功能表型选择可用时，我们就可以勾勒出蛋白质的结构和进化景观，而无需使用纯化的蛋白质、收集物理测量结果或依赖天然蛋白质变体库。