CAREER: Lipid Regulation of Receptor Tyrosine Kinases

有了这个奖项，化学部的生命过程化学计划正在资助亚当W。Smith博士研究了受体酪氨酸激酶的脂质调节。活细胞的表面由嵌入有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是功能不可或缺的，但解决这些相互作用已被证明是极具挑战性的。该项目应用先进的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。长期目标是建立一个定量化学模型，用于质膜脂质和一类称为受体酪氨酸激酶（RTK）的膜蛋白之间的界面。RTK是调节细胞生长和分化的整合膜蛋白。该项目的重点是两个RTK，EphA 2和EGFR。与这些研究目标不可分割的是一个教育和更广泛的影响计划，提高阿克伦大学的STEM教育，并为全国各地的实验室教学提供材料和课程。这包括开发3D可打印的智能手机光谱仪（SpecPhone），用于在大学实验室课程中实施。SpecPhone的低成本和简单性也使K-12学生和公民能够使用它，以便他们能够参与现实世界的科学问题。 更广泛的影响工作创建新的课程和教师培训研讨会的K-12 STEM教育，参加当地的制造商博览会，并制定协议的一个变革性的公民科学项目在伊利湖流域。脂质溶剂化膜蛋白，并在许多情况下，通过直接，具体的接触，调节其活动。许多脂质-蛋白质相互作用是从静态结构或计算机模拟中推断出来的;然而，很少有实验数据来验证这些相互作用并确定其动力学和热力学稳定性。该项目使用先进的荧光方法，包括脉冲交错激发荧光互相关光谱（PIE-FCCS）和单分子成像来测量生物膜中的分子缔合。长期的目标是建立一个定量的化学模型，质膜脂质和受体酪氨酸激酶（RTK）之间的接口。RTK的胞外结构域（ECD）和胞内结构域（ICD）都直接与质膜结合，但这种结合的化学细节还不清楚。该项目重点关注两种RTK，EphA 2和EGFR。核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。为了验证这一假设，我们确定了模型支持的脂质双层中阴离子脂质结合EGFR的亲和力和特异性。我们还研究了模型膜中阴离子脂质对EphA 2结构和动力学的调节。最后，我们解决了阴离子脂质结合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，头基结构，溶剂pH值和盐效应等参数的影响。这些结果大大推进了我们对指导细胞通讯的化学相互作用的理解。与这些研究目标不可分割的是一个教育和更广泛的影响计划，提高阿克伦大学的STEM教育，并为全国各地的实验室教学提供材料和课程。这包括开发3D可打印的智能手机光谱仪（SpecPhone），用于在大学实验室课程中实施。SpecPhone的低成本和简单性也使K-12学生和公民能够使用它，以便他们能够参与现实世界的科学问题。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研讨会，参加当地的创客博览会，并为伊利湖流域的变革性公民科学项目制定协议。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。

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.