CAREER: Structural dynamics of post-translationally modified Calmodulin and its role in target recognition

细胞通过产生、传递和接收化学信号进行交流。钙调蛋白存在于所有高等生物体中，它能感知细胞中的钙水平，调节重要功能，如心跳、肌肉收缩、学习和记忆。该项目旨在确定允许这种蛋白质调节这些关键功能的基本原理。有趣的是，钙调素可以激活或禁用这些功能，但钙调素选择这种作用的过程尚不清楚。该项目将研究可能改变蛋白质柔韧性及其三维结构的蛋白质修饰。获得的知识将有助于解释某些修饰如何调节钙调素与其靶标相互作用的方式。该项目的更广泛影响包括一个全面的教育和培训计划，以准备一个高素质的本科生，研究生和博士后研究人员在各个学科的池。特别是，该计划将创建丰富的课程项目，提供使用最先进仪器的实践经验的新机会，并通过创造新的出国留学机会培养具有全球竞争力的劳动力。为了提高普通民众的科学素养，克莱姆森大学将建立一个新的视频图书馆。该项目的目标是了解钙调蛋白的结构、动力学和磷酸化状态如何通过提供所需的靶标特异性和选择性来调节靶标识别。该项目将采用综合的方法，结合荧光光谱工具、生物物理方法和分子动力学模拟来表征钙调素在不同磷酸化状态下的三维结构和动力学。为了实现这一目标，该计划将分为三个不同的任务。第一项任务将涉及生成野生型钙调素的结构模型，并使用基于Förster共振能量转移实验的混合结构动态工具将野生型钙调素与各种磷酸化状态进行比较。第二项任务是确定野生型钙调蛋白的中央螺旋对不同磷酸化状态的动力学差异。第三个也是最后一个任务是确定钙调素磷酸化状态对目标识别的不同影响。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Cells communicate by generating, transmitting and receiving chemical signals. The protein Calmodulin, present in all higher organisms, senses calcium levels in the cell and regulates vital functions such as heart beating, muscle contraction, learning and memory. This project is geared towards identifying the fundamental principles that allow this protein to regulate these critical functions. Interestingly, Calmodulin can activate or deactivate these functions, but the process by which Calmodulin selects this action is not understood. The project will study modifications to the protein that are likely to alter the protein's flexibility and its three-dimensional structure. The obtained knowledge would help explain how certain modifications regulate the way Calmodulin interacts with its targets. The broader impacts of this project include the incorporation of a thorough educational and training program to prepare a pool of highly qualified undergraduate, graduate, and postdoctoral researchers across various disciplines. In particular, the program will create enriched curricular programs that provide new opportunities for hands-on experience using state-of-the-art instrumentation and train a globally competent workforce by creating new study abroad opportunities. To improve scientific literacy among the general population, a new video library is to be broadcasted from Clemson University.The goal of the project is to understand how the structure, dynamics and phosphorylation state of Calmodulin modulate target recognition by providing the required target specificity and selectivity. The PI will use an integrative approach combining fluorescence spectroscopic tools, biophysical methods, and molecular dynamic simulations to characterize the three-dimensional structure and the dynamics of Calmodulin at different phosphorylated states. To accomplish this goal, the program will be split into three distinct tasks. The first task will involve the generation of structural models of wild-type Calmodulin and the comparison of the wild-type to various phosphorylated states using a hybrid structural dynamic tool based on Förster resonance energy transfer experiments. The second task is to determine the difference in the dynamics of the central helix of the wild-type Calmodulin against various phosphorylated states. The third and final task is to determine the differential impact of the phosphorylation state of Calmodulin on target recognition.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.