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

由生物技术、生物化学和生物质工程项目以及生物传感项目资助的这一创新和高风险的NSF EAGER奖的总体目标是使用定向进化来修饰新型钙调节的β - roll肽，使它们能够与具有内置变构识别的蛋白质靶标结合。卷曲肽具有软木塞螺旋结构，在钙存在下可逆形成。这个结构域还没有进化到与其他蛋白质结合，但在结构上它类似于富含亮氨酸的重复序列和锚蛋白重复序列，这两种蛋白质都经过进化和改造，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β - roll肽，这些肽可以与模型蛋白靶点结合，并且支架的内置变弹性调节将使结合界面能够根据钙浓度形成。这将是生物分子识别的内在无序肽的定向进化的第一个例子。这些肽可以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.