CAREER: Tyrosine Phosphorylation and Kinase Activity as a Regulator of Sodium-Glucose Transporters

职业：酪氨酸磷酸化和激酶活性作为钠-葡萄糖转运蛋白的调节剂

• 批准号: 2340201
• 负责人: Jason Sprowl
• 金额: $ 74.69万
• 依托单位: SUNY at Buffalo
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-03-01 至 2029-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2340201&HistoricalAwards=false
• 关键词: CAREER; Tyrosine; Phosphorylation; Kinase; Activity

The sodium-glucose-linked transporters (SGLTs) are proteins that are necessary for cellular uptake of glucose, which is an essential nutrient. Altered activity of these proteins can lead to poor glucose absorption and nutrient deficiency. Unfortunately, cellular events that regulate SGLT activity are poorly understood. This is particularly true for tyrosine phosphorylation, a form of modification that can change protein structure and function, that has not yet been studied in relation to SGLT activity. Therefore, the long-term goal of this NSF CAREER research project is to improve the understanding of how SGLT tyrosine phosphorylation, or tyrosine kinase proteins that activate this form of modification, regulate changes in glucose movement into cells. The project also includes several strategies for educational improvements that include highlighting the biological importance of SGLTs, as well as the training and recruitment of junior scientists who will lead future science, technology, engineering, and mathematic efforts. Collectively, this research project is expected to impact many scientific disciplines, including molecular, cellular, and systems biology, through educational insight and novel scientific discoveries involving glucose transporters.The research objective of this NSF CAREER project is to identify a novel post-translational event that regulates variability in SGLT activity and glucose absorption. Solute carrier transporters, including SGLTs, have been associated as passive pores that move substrates across cell membranes alone or with electrochemical gradients. However, the hypothesis to be tested as part of the current project is that SGLTs are tyrosine phosphorylated by specific kinases at the cell surface, and that these phosphorylation events are associated with transport function. To address this hypothesis, the project will 1) evaluate the phosphorylation status of SGLTs in different cellular compartments using cell fractionation and mass spectrometry; 2) identify the role of tyrosine phosphorylation sites in regulating SGLT activity using site-directed mutagenesis, kinase inhibitors, and SGLT activity assays; and 3) identify tyrosine kinases that mediate SGLT activity using phosphoproteomic analysis and kinase interaction assays. Additionally, the strategies and outcomes from the proposed research objectives will be used to improve basic scientific knowledge, generate a passion for research, and improve leadership capabilities in the field of biological sciences. This will be done by establishing an annual summer research position for underprivileged high school students, working with middle school educators to increase recognition of reproducible and high-quality science, and developing online content that will increase familiarity with transporter proteins.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.

葡萄糖连接转运蛋白（SGLT）是细胞摄取葡萄糖所必需的蛋白质，这是必不可少的养分。这些蛋白质的活性改变会导致葡萄糖吸收和营养缺乏症。不幸的是，调节SGLT活性的细胞事件知之甚少。对于酪氨酸磷酸化尤其如此，这是一种可以改变蛋白质结构和功能的修饰形式，尚未与SGLT活性有关。因此，该NSF职业研究项目的长期目标是提高人们对激活这种修饰形式的SGLT酪氨酸磷酸化或酪氨酸激酶蛋白如何调节葡萄糖运动向细胞的变化的理解。该项目还包括几种教育改进的策略，包括强调SGLT的生物学重要性，以及将领导未来科学，技术，工程和数学努力的初级科学家的培训和招聘。总的来说，该研究项目有望通过教育洞察力和涉及葡萄糖转运蛋白的新科学发现，包括分子，细胞和系统生物学，包括分子，细胞和系统生物学。该NSF职业项目的研究目标是确定一种新型的翻译后事件，该事件调节了SGLT活性和吸收SGLT活性和gl糖吸收的可变性。包括SGLT在内的溶质载体转运蛋白已成为被动孔，它们单独或与电化学梯度一起移动底物。然而，作为当前项目的一部分进行检验的假设是，SGLT是由细胞表面的特定激酶磷酸化的酪氨酸，并且这些磷酸化事件与运输功能有关。为了解决这一假设，该项目将使用细胞分馏和质谱法评估不同细胞室中SGLT的磷酸化状态； 2）确定酪氨酸磷酸化位点在使用位置定向的诱变，激酶抑制剂和SGLT活性测定中调节SGLT活性中的作用； 3）鉴定使用磷酸蛋白质组学分析和激酶相互作用测定法介导SGLT活性的酪氨酸激酶。此外，提议的研究目标的策略和结果将用于改善基本科学知识，对研究产生热情，并提高生物科学领域的领导能力。 This will be done by establishing an annual summer research position for underprivileged high school students, working with middle school educators to increase recognition of reproducible and high-quality science, and developing online content that will increase familiarity with transporter proteins.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.