Design of switchable proteins and enzymes.

 描述(由申请人提供)：该项目采用实验和计算相结合的方法来开发普通蛋白质可以转化为分子开关的机制。诱导结构域交换(INDOS)是一种允许蛋白质或酶的功能通过小分子药物、金属离子或pH变化来开启和关闭的技术。INDOS将被用来在活细胞中实时操纵细胞通路，以检查它们在健康和疾病中的作用。蛋白质片段交换(FREX)和交替框架折叠(AFF)是将任意结合蛋白转变为荧光生物传感器的机制。我们将生产一系列传感器，用于检测体内和体外的各种生物靶标。这项研究的影响在于，正在开发的开关机制：(I)是通用的，可以应用于许多蛋白质；(Ii)是通用的，允许所产生的开关执行各种新的和有用的功能；以及(Iii)将提供对其潜在的分子现象(结构域交换、片段互补和循环排列)的洞察，所有这些都在自然界中普遍存在，但人们对这些现象知之甚少。

 DESCRIPTION (provided by applicant): This project employs a combined experimental and computational approach to develop mechanisms by which ordinary proteins can be converted into molecular switches. Induced domain swapping (INDOS) is a technology that allows the function of a protein or enzyme to be switched on and off by a small molecule drug, metal ion, or change in pH. INDOS will be used to manipulate cellular pathways, in living cells and in real time, to examine their roles in health and disease. Protein fragment exchange (FREX) and alternate frame folding (AFF) are mechanisms by which an arbitrary binding protein can be turned into a fluorescent biosensor. We will generate a family of sensors for detecting a variety of biological targets in vivo and in vitro. The impact of this study lies in that the switching mechanisms being developed: (i) are general and can be applied to many proteins; (ii) are versatile, allowing the resulting switches to perform a variety of new and useful functions; and (iii) will provide insight into their underlying molecular phenomena (domain swapping, fragment complementation, and circular permutation), all of which occur commonly in nature yet are poorly understood.