CAREER: Lipid Regulation of Receptor Tyrosine Kinases

职业：受体酪氨酸激酶的脂质调节

• 批准号: 2308307
• 负责人: Adam Smith
• 金额: $ 65万
• 依托单位: Texas Tech University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-15 至 2024-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2308307&HistoricalAwards=false
• 关键词: CAREER; Lipid; Regulation; Receptor; Tyrosine

With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Adam W. Smith from the University of Akron to investigate lipid regulation of receptor tyrosine kinases. The surface of living cells is composed of a lipid membrane embedded with thousands of protein receptors. Dynamic associations between membrane proteins and lipids are integral to function, but resolving these interactions has proven to be extremely challenging. This project applies advanced fluorescence methods, and single-molecule imaging to measure lipid-protein interactions in biological membranes. The long-term goal is to develop a quantitative chemical model for the interface between plasma membrane lipids and a class of membrane proteins called receptor tyrosine kinases (RTKs). RTKs are integral membrane proteins that regulate cell growth and differentiation. The focus of this project is on two RTKs, EphA2 and EGFR. Integral with these research objectives is an education and broader impacts program that enhances STEM education at the University of Akron and provides materials and curriculum for enhanced laboratory instruction across the country. This includes the development of a 3D-printable, smartphone spectrometer (the SpecPhone) for implementation in university laboratory courses. The low cost and simplicity of the SpecPhone also make it accessible to K-12 students and citizens so that they can engage in real-world science problems. The broader impacts work creates new curriculum and teacher training workshops for K-12 STEM education, to participate in local Maker Fairs, and to develop protocols for a transformative citizen science project in the Lake Erie watershed.Lipids solvate membrane proteins and, in many cases, regulate their activity through direct, specific contacts. Many lipid-protein interactions are inferred from static structures or computer simulations; however, there is little experimental data to verify these interactions in situ and determine their kinetic and thermodynamic stability. This project uses advanced fluorescence methods, including pulsed-interleaved excitation fluorescence cross-correlation spectroscopy (PIE-FCCS) and single-molecule imaging to measure molecular associations in biological membranes. The long-term goal is to develop a quantitative chemical model for the interface between plasma membrane lipids and receptor tyrosine kinases (RTKs). Both the extracellular domain (ECD) and the intracellular domain (ICD) of RTKs associate directly with the plasma membrane, but the chemical details of the associations are not well-understood. This project focuses on two RTKs, EphA2 and EGFR. The central hypothesis is that anionic lipids bind these proteins and regulate their structure and activity. To test this hypothesis, we determine the affinity and specificity of anionic lipid binding to EGFR in model supported lipid bilayers. We also investigate the regulation of EphA2 structure and dynamics by anionic lipids in model membranes. Finally, we resolve the functional role of anionic lipid binding to EGFR. Achieving these objectives contributes to a systematic understanding of how the lipid-protein interface is affected by parameters like lipid charge, headgroup structure, solvent pH, and salt effects. The results significantly advance our understanding of the chemical interactions that guide cell communication. Integral with these research objectives is an education and broader impacts program that enhances STEM education at the University of Akron and provides materials and curriculum for enhanced laboratory instruction across the country. This includes the development of a 3D-printable, smartphone spectrometer (the SpecPhone) for implementation in university laboratory courses. The low cost and simplicity of the SpecPhone also make it accessible to K-12 students and citizens so that they can engage in real-world science problems. The focus of the broader impacts work is to create new curriculum and teacher training workshops for K-12 STEM education, to participate in local Maker Fairs, and to develop protocols for a transformative citizen science project in the Lake Erie watershed.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.

通过此奖项，化学生命过程的化学过程计划正在为阿克伦大学的亚当·W·史密斯博士提供资金，以调查受体酪氨酸激酶的脂质调节。活细胞的表面由嵌入数千种蛋白质受体的脂质膜组成。膜蛋白和脂质之间的动态关联对于功能是不可或缺的，但是解决这些相互作用已被证明是极具挑战性的。该项目采用晚期荧光方法和单分子成像来测量生物膜中的脂质 - 蛋白质相互作用。长期目标是为质膜脂质与一类称为受体酪氨酸激酶（RTKS）的膜蛋白之间的界面开发定量化学模型。 RTK是调节细胞生长和分化的整体膜蛋白。该项目的重点是两个RTK，EPHA2和EGFR。与这些研究目标不可或缺的是一项教育和更广泛的影响计划，该计划可以增强阿克伦大学的STEM教育，并提供材料和课程，以增强全国的实验室教学。这包括开发3D打印机的智能手机光谱仪（Specphone），用于在大学实验室课程中实施。特定手机的低成本和简单性也使K-12学生和公民可以使用它，以便他们可以涉及现实的科学问题。 更广泛的影响工作为K-12 STEM教育创建了新的课程和教师培训研讨会，以参加本地制造商博览会，并为伊利湖水域中的变革性公民科学项目制定协议。lipids溶解膜膜蛋白，并在许多情况下通过直接的，具体的，具体的接触来调节其活动。从静态结构或计算机模拟推断出许多脂质 - 蛋白质相互作用。但是，几乎没有实验数据来验证这些相互作用的原位并确定它们的动力学稳定性。该项目使用晚期荧光方法，包括脉冲中断激发荧光互相关光谱（PIE-FCC）和单分子成像以测量生物膜中的分子相关性。长期目标是为质膜脂质和受体酪氨酸激酶（RTKS）之间的界面开发定量化学模型。 RTK的细胞外结构域（ECD）和细胞内结构域（ICD）均直接与质膜相关，但是关联的化学细节并不理解。该项目侧重于两个RTK，EPHA2和EGFR。中心假设是阴离子脂质结合这些蛋白质并调节其结构和活性。为了检验这一假设，我们确定了在模型支持的脂质双层中与EGFR结合的阴离子脂质结合的亲和力和特异性。我们还研究了模型膜中阴离子脂质对EPHA2结构和动力学的调节。最后，我们解决了阴离子脂质与EGFR的功能作用。实现这些目标有助于系统地了解脂质 - 蛋白质界面如何受到脂质电荷，头部组结构，溶剂pH和盐效应等参数的影响。结果大大提高了我们对指导细胞通信的化学相互作用的理解。与这些研究目标不可或缺的是一项教育和更广泛的影响计划，该计划可以增强阿克伦大学的STEM教育，并提供材料和课程，以增强全国的实验室教学。这包括开发3D打印机的智能手机光谱仪（Specphone），用于在大学实验室课程中实施。特定手机的低成本和简单性也使K-12学生和公民可以使用它，以便他们可以涉及现实的科学问题。更广泛的影响工作的重点是为K-12 STEM教育创建新的课程和教师培训讲习班，参加本地制造商博览会，并为伊利湖水上的变革性公民科学项目制定协议。这项奖项反映了NSF的法定任务，并通过使用基金会的智力效果和宽阔的范围来评估NSF的法定任务，并值得通过评估来进行评估。

项目成果

Direct Quantification of Serum Protein Interactions with PEGylated Micelle Nanocarriers

血清蛋白与聚乙二醇化胶束纳米载体相互作用的直接定量

• DOI: 10.1021/acs.biomac.2c01538
• 发表时间: 2023
• 期刊: Biomacromolecules
• 影响因子: 6.2
• 作者: Mallory, D. Paul;Freedman, Abegel;Kaliszewski, Megan J.;Montenegro-Galindo, Gladys Rocío;Pugh, Coleen;Smith, Adam W.
• 通讯作者: Smith, Adam W.