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CAREER: Lipid Regulation of Receptor Tyrosine Kinases

职业：受体酪氨酸激酶的脂质调节

基本信息

批准号：
1753060
负责人：
Adam Smith
金额：
$ 65万
依托单位：
University of Akron
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2018
资助国家：
美国
起止时间：
2018-06-01 至 2022-12-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1753060&HistoricalAwards=false
关键词：
CAREER Lipid Regulation Receptor Tyrosine

项目摘要

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.

有了这个奖项，化学部门的生命过程化学项目将资助阿克伦大学的亚当·w·史密斯博士研究受体酪氨酸激酶的脂质调节。活细胞的表面是由一层脂质膜组成的，脂质膜上嵌有成千上万的蛋白质受体。膜蛋白和脂质之间的动态关联是功能的组成部分，但解决这些相互作用已被证明是极具挑战性的。本项目应用先进的荧光方法和单分子成像技术来测量生物膜中脂质-蛋白的相互作用。长期目标是开发质膜脂和一类称为受体酪氨酸激酶（RTKs）的膜蛋白之间界面的定量化学模型。RTKs是调节细胞生长和分化的完整膜蛋白。这个项目的重点是两个rtk， EphA2和EGFR。与这些研究目标相结合的是一个教育和更广泛的影响项目，该项目旨在加强阿克伦大学的STEM教育，并为全国范围内的实验室教学提供材料和课程。这包括开发一种可3d打印的智能手机光谱仪（SpecPhone），用于大学实验室课程的实施。SpecPhone的低成本和简单性也使K-12学生和市民可以使用它，这样他们就可以参与现实世界的科学问题。更广泛的影响工作为K-12 STEM教育创造了新的课程和教师培训讲习班，参加了当地的创客博览会，并为伊利湖流域的一个变革性公民科学项目制定了协议。脂质溶剂化膜蛋白，在许多情况下，通过直接的、特定的接触调节膜蛋白的活性。许多脂质与蛋白质的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，并确定它们的动力学和热力学稳定性。本项目采用先进的荧光方法，包括脉冲交错激发荧光相互关联光谱（PIE-FCCS）和单分子成像来测量生物膜中的分子关联。长期目标是建立质膜脂和受体酪氨酸激酶（RTKs）之间界面的定量化学模型。RTKs的胞外结构域（ECD）和胞内结构域（ICD）都与质膜直接相关，但这种关联的化学细节尚不清楚。本项目重点研究EphA2和EGFR两种rtk。核心假设是阴离子脂质结合这些蛋白质并调节它们的结构和活性。为了验证这一假设，我们在模型支持的脂质双分子层中确定了阴离子脂质与EGFR结合的亲和力和特异性。我们还研究了阴离子脂质在模型膜中对EphA2结构和动力学的调节。最后，我们分析了阴离子脂质结合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和盐效应等参数的影响。这些结果极大地促进了我们对指导细胞通讯的化学相互作用的理解。与这些研究目标相结合的是一个教育和更广泛的影响项目，该项目旨在加强阿克伦大学的STEM教育，并为全国范围内的实验室教学提供材料和课程。这包括开发一种可3d打印的智能手机光谱仪（SpecPhone），用于大学实验室课程的实施。SpecPhone的低成本和简单性也使K-12学生和市民可以使用它，这样他们就可以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，并为伊利湖流域的一个变革性公民科学项目制定协议。该奖项反映了美国国家科学基金会的法定使命，并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，被认为值得支持。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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.

A 3D-Printable Dual Beam Spectrophotometer with Multiplatform Smartphone Adaptor