Research Initiation Award: Redox Regulated Phosphorylation of BCAT by PKC

研究启动奖：PKC 氧化还原调节 BCAT 磷酸化

• 批准号: 2401097
• 负责人: Ming Dong
• 金额: $ 29.94万
• 依托单位: University of North Carolina at Wilmington
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-10-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2401097&HistoricalAwards=false
• 关键词: Research; Initiation; Award; Redox; Regulated

Research Initiation Awards provide support for junior and mid-career faculty at Historically Black Colleges and Universities who are building new research programs or redirecting and rebuilding existing research programs. It is expected that the award helps to further the faculty member's research capability and effectiveness, improve research and teaching at the home institution, and involves undergraduate students in research experiences. The award to North Carolina Agricultural & Technical State University has potential to increase the understanding of the cell membrane. The proposed study intends to train undergraduate and graduate students in the study of an enzyme that affects cell membrane function. The PI will actively engage and recruit underrepresented minority students into the project so that the students may engage in cutting edge research. This activity will enhance the technical and critical thinking skills of students.The goal of this proposal is to study the PKCα-BCATc interaction via biochemical and biophysical methods. Previously the PI showed that the membrane localization of cytosolic human Branched-Chain Aminotransferase (BCATc) regulates the autophagosome synthesis which further regulates Alzheimer’s Disease (AD), and the membrane localization of BCATc is redox regulated by the Protein Kinase C α (PKCα) mediated phosphorylation. In this proposed work, the PI will identify the PKCα-mediated phosphorylation site in BCATc, express and purify the PKCα-phosphorylated BCATc, and solve the crystal structure of the phosphorylated BCATc to analyze the structural changes of BCATc triggered by the phosphorylation by PKCα. The work will analyze the structure of PKCα-BCATc complex via co-crystallization and other biophysical methods, e.g. SPRi to uncover the protein-protein interaction interface and further understand the mechanism of the phosphorylation process. The constructed model built from the project outcomes will enable the PI to understand the mechanism of how the redox-regulated phosphorylation of BCATc by PKCα regulates its conformational changes that further regulate its membrane localization. This is the first time that these interactions will be described at the molecular level, and this would be a significant contribution to the field and help advance the understanding of these two inter-related modes of BCATc regulation.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.

研究启动奖为历史上黑人学院和大学的初级和中级职业教师提供支持，他们正在建立新的研究项目或重新定向和重建现有的研究项目。预计该奖项将有助于进一步提高教师的研究能力和效率，改善家庭机构的研究和教学，并使本科生参与研究经验。授予北卡罗来纳州农业技术州立大学的奖项有可能增加对细胞膜的理解。这项拟议的研究旨在培养本科生和研究生研究影响细胞膜功能的酶。PI将积极参与并招募代表性不足的少数民族学生参加该项目，以便学生可以从事尖端研究。这项活动将提高学生的技术和批判性思维能力。本计划的目标是通过生物化学和生物物理方法研究PKCα-BCATc相互作用。先前PI显示胞质人类支链氨基转移酶（BCATc）的膜定位调节自噬体的合成，自噬体的合成进一步调节阿尔茨海默病（AD），并且BCATc的膜定位由蛋白激酶C α（PKCα）介导的磷酸化进行氧化还原调节。在本研究中，PI将鉴定BCATc中PKCα介导的磷酸化位点，表达和纯化PKCα磷酸化的BCATc，并解析磷酸化的BCATc的晶体结构，以分析PKCα磷酸化引发的BCATc结构变化。本工作将通过共结晶和其他生物物理方法，如SPRi，分析PKCα-BCATc复合物的结构，以揭示蛋白质-蛋白质相互作用界面，并进一步了解磷酸化过程的机制。根据项目成果构建的模型将使PI能够理解PKCα如何通过氧化还原调节BCATc的磷酸化来调节其构象变化，从而进一步调节其膜定位的机制。这是第一次在分子水平上描述这些相互作用，这将是对该领域的重大贡献，并有助于促进对BCATc调节的这两种相互关联模式的理解。该奖项反映了NSF的法定使命，并被认为值得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来支持。