Control of Protein Dimerization through Light-Regulated Rapamycin

凭借这一奖项，化学部生命过程化学项目正在资助匹兹堡大学的亚历山大·迪特斯开发一种新型分子“胶水”，可用于将两个蛋白质分子结合在一起，从而控制多种生物功能。在这个研究项目中开发的新型“胶水”类似于一种被称为雷帕霉素的天然物质，这种物质已被广泛用于结合蛋白质对。这种新物质与雷帕霉素不同，因为它有一个光敏开关。这个开关允许研究人员随意打开和关闭蛋白质结合过程，产生一个强大的新工具。这项研究开辟了各种可能的新应用，可能对许多生物和医学研究领域产生广泛影响。研究人员正在与一家儿童博物馆合作，进一步扩大他们工作的影响，开发演示和展览，为幼儿和他们的父母带来基本的生物学事件，特别是那些涉及紫外线与组织相互作用的事件。在这个项目中，研究人员正在开发光响应雷帕霉素类似物，用于蛋白质二聚化和其他细胞过程的光学控制。雷帕霉素是一种常用的生物分子工具，已被用于控制广泛的生理过程。许多细胞功能都在其控制之下，包括转录、信号转导、蛋白定位、酶激活、蛋白剪接、蛋白分泌和细胞中的蛋白翻译后修饰。雷帕霉素通过募集FKBP和FRB以形成稳定的三元复合物而作为蛋白质二聚化的小分子诱导剂。通过设计和合成雷帕霉素的光敏类似物，正在开发一种适用于许多生物学应用的通用且普遍适用的光调节方法。光代表了一种创新的外部控制元件，因为a）它与绝大多数细胞过程正交，B）它是微创的，以及c）它可以以非常高的空间和时间分辨率进行控制。雷帕霉素的光激活类似物与融合至FKBP和FRB的不同蛋白质结合，将使得能够光学调节广泛的细胞过程，包括蛋白质功能、蛋白质定位和蛋白质缔合。研究目标的完成将导致1）正交笼状雷帕霉素-FKBP对的开发，用于多个细胞过程的多波长激活，以及2）通过用光开关基序修饰的雷帕霉素类似物实现生物过程的可逆光开关。所提出的研究的预期成果是各种细胞过程的时空控制的统一方法。

With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Alexander Deiters of the University of Pittsburgh to develop a new type of molecular "glue" that can be used to bind together two molecules of protein and, thus, control a variety of biological functions. The new type of "glue" being developed in this research project is similar to a naturally occurring substance known as rapamycin that has been extensively used to bind pairs of proteins. The new substance differs from rapamycin since it has a light-sensitive switch. This switch allows the investigators to turn on and off the protein-binding process at will, producing a powerful new tool. This research is opening up a wide variety of possible new applications that would potentially have a broad impact on a number of biological and medical fields of study. The investigators are further broadening the impact of their work by partnering with a children's museum to develop demonstrations and exhibits that bring basic biological events, especially those involving the interaction of ultraviolet light with tissues, to life for young children and their parents.In this project, the investigators are developing light-responsive rapamycin analogs for the optical control of protein dimerization and other cellular processes. Rapamycin is a commonly applied biomolecular tool that has been used to control a wide range of physiological processes. An extensive number of cellular functions have been brought under its control, including transcription, signal transduction, protein localization, enzyme activation, protein splicing, protein secretion, and protein post-translational modification in cells. Rapamycin functions as a small molecule inducer of protein dimerization by recruiting FKBP and FRB to form a stable ternary complex. A versatile and generally applicable light-regulation methodology that is adaptable to numerous biological applications is being developed by designing and synthesizing light-sensitive analogs of rapamycin. Light represents an innovative external control element, since a) it is orthogonal to the vast majority of cellular processes, b) it is minimally invasive, and c) it can be controlled with very high spatial and temporal resolution. Light-activated analogs of rapamycin, in conjunction with different proteins fused to FKBP and FRB, will enable the optical regulation of a wide range of cellular processes, including protein function, protein localization, and protein association. Completion of the research objectives will lead to 1) the development of orthogonal caged rapamycin-FKBP pairs for the multi-wavelength activation of multiple cellular processes, and 2) the reversible light-switching of biological processes through rapamycin analogs modified with photo-switchable motifs. Expected outcomes of the proposed research are a unifying approach for the spatio-temporal control of various cellular processes.