EAGER: Evolution of Beta Roll Peptides to Create Allosterically-Regulated Binding Domains Using Bacterial Cell Surface Display

1161160/Banta由生物技术、生化和生物质工程计划和生物传感计划资助的这一创新和高风险的国家科学基金会EIGER奖的总体目标是利用定向进化来修饰新型的钙调节贝塔卷曲多肽，使它们能够通过内置变构识别与蛋白质靶标结合。贝塔卷肽具有软木塞螺旋结构，在有钙存在时可逆形成。这个结构域还没有进化到与其他蛋白质结合，但在结构上类似于富含亮氨酸的重复蛋白和锚蛋白重复蛋白，这两个重复蛋白都已经进化并被设计用于生物分子识别。研究人员已经开始使用定向进化来创造新的与模型蛋白质靶标结合的贝塔卷肽，支架的内置变构调节将使结合界面的形成取决于钙浓度。这将是用于生物分子识别的内在无序多肽定向进化的第一个例子。这些多肽随后可用于传感或其他分子工程应用。

1161160/ BantaThe overall goal of this innovative and high risk NSF EAGER award, funded by the Biotechnology, Biochemical and Biomass Engineering Program and by the Biosensing Program, is to use directed evolution to modify novel calcium-regulated beta roll peptides so that they can bind to protein targets with built-in allosteric recognition. The beta roll peptide has a cork-screw structure that reversibly forms in the presence of calcium. This domain has not evolved to bind to other proteins, but structurally it resembles the leucine rich repeat and ankyrin repeat proteins, which have both evolved, and been engineered, for biomolecular recognition. The researchers have begun to use directed evolution to create new beta roll peptides that bind to a model protein target, and the built-in allosteric regulation of the scaffold will enable the binding interface to be formed depending on the calcium concentration. This will be the first example of the directed evolution of an intrinsically disordered peptide for biomolecular recognition. These peptides can then be used in sensing or other molecular engineering applications.