The underlying dynamic exchange that dictates serine protease function

决定丝氨酸蛋白酶功能的潜在动态交换

• 批准号: 2332239
• 负责人: Elan Eisenmesser
• 金额: $ 68.76万
• 依托单位: University of Colorado at Denver
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-02-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2332239&HistoricalAwards=false
• 关键词: underlying; dynamic; exchange; dictates; serine

Proteases comprise a large group of enzymes responsible for cleaving (or processing) a wide array of substrates. One class of proteases, called serine proteases, are predicted to move between inactive and active conformations. These movements are critical for serine proteases to function and represent one of the clearest examples of why the ability of an enzyme to move is so important for their functions. In this research, a single site within the serine protease called exfoliative toxin (ETA) has been identified that moves this enzyme either completely to its inactive or active conformation. By changing this single site through mutagenesis of the natural amino acid, the explicit inactive and active structures of ETA can be determined. Further methods will also be used to understand how different parts of ETA communicate this conformational change between inactive and active conformations to different parts of the enzyme, referred to generally as “allostery”. This project will train members of the underrepresented groups through multiple programs, including the Graduate Experience for Multicultural Students (GEMS) and a new program through the School of Medicine called Colorado Research Experiences (CORE).This research will capitalize on the novel identification of a single inherently dynamic residue, ETA D164, situated adjacent to the active site. This residue acts as a pivotal “switch” that governs the inherent global sampling of inactive and active conformations, as proposed for serine proteases over a decade ago. Through mutagenesis, shifts in solution ensembles are observed as monitored via chemical shift perturbations (CSPs) in either of two directions which correlates to activity to shift in either of two directions. Thus, the goals in this research are to 1) Identify the allosteric networks coupled within ETA that modulate global sampling of inactive and active conformations and 2) determine both the dynamic and structural basis of this sampling of inactive and active conformations. This project is supported by the Molecular Biophysics Cluster of the Division of Molecular and Cellular Biosciences.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.

蛋白酶包含一组负责裂解（或加工）的酶，现在是不活跃的构型的。已经确定该酶通过更改单个单个单个单个单个单个单个单个单一的表现，可以确定酶的构象与多个程序的差异。学生（宝石）和一个新课程Olorado研究经验（Core）。这项研究将利用单个固有动态残留物的新颖识别，该残留物是该残留物的构象。丝网在十年前，在解决方案中的三个方向中的任何一个方向都可以通过两个方向纠正了两个方向的活动。在ETA内进行变构，以调节活性构象的全局采样2）确定无效和主动置信度采样的动态和结构基础。通过基金会的智力优点和更广泛的影响审查标准评估，值得一提。