Mechanistic studies of membrane lateral organization in cell plasma membranes.

描述(申请人提供)：我提出的研究的目标是描述细胞质膜中脂质介导的横向组织的特征，并破译导致这种结构的潜在物理机制。在这样做的过程中，我将通过开发新的实验工具来操纵和测试活细胞中脂质介导的组织的功能后果，从而克服目前细胞膜研究面临的重大障碍。我过去在破译简单模型膜中的液体不混溶方面的成功，为我目前和未来在生物膜方面的博士后研究奠定了基础。自从加入康奈尔大学的贝尔德实验室以来，我发现了从RBL肥大细胞分离出的质膜囊泡的强烈临界波动。这一结果有力地支持了我的工作假设，即在生理温度下细胞质膜存在临界波动。在这个奖项的指导阶段，我将开始直接测试我的工作假设，通过实施实验和分析技术来量化和解释完整细胞表面的纳米级组织(Aim1)。1还将开发实验方法，以调节质膜囊泡和完整细胞质膜的横向异质性(目标2)。通过这些学习，我将获得必要的技能和观点，成功地将我强大的身体背景应用于这一与生物医学相关的研究领域。作为一名独立的研究人员，我将继续这两个目标的工作，最终破译膜横向组织(Aim1)的物理基础，并开发新的方法来探索脂质介导的横向异质性在RBL肥大细胞中IgE介导的信号转导中的功能后果(Aim 2)。我的长期目标是开发通用的方法来探索脂类在广泛的过程和细胞类型中的功能作用，这将使我们能够对脂类介导的细胞质膜组织的生物医学后果进行新的研究。独立之路奖将给我提供资源，让我在博士后时期深入关注我的生物培训，并在我过渡到未来作为一名年轻教员的职位时，独立追求自己的研究目标。 相关性：人们普遍认为质膜的横向异质性在正常的细胞过程以及人类疾病的发展和维持中起着重要的功能作用。我将通过开发有效的实验技术和一个清晰的概念框架来研究天然膜中的脂质和蛋白质，从而为干预基于细胞膜的病理学开辟新的机会。

DESCRIPTION (provided by applicant): The goal of my proposed research is to characterize lipid-mediated lateral organization in cell plasma membranes and to decipher the underlying physical mechanisms that give rise to this structure. In so doing, I will surmount significant barriers currently facing cell membrane research by developing new experimental tools to manipulate and test the functional consequences of lipid-mediated organization in living cells. My past success in deciphering liquid immiscibility in simple model membranes forms the foundation for my current and future postdoctoral studies in biological membranes. Since joining the Baird Laboratory at Cornell University, I have identified robust critical fluctuations in plasma membrane vesicles isolated from RBL mast cells. This result strongly supports my working hypothesis that critical fluctuations are present in cell plasma membranes at physiological temperatures. In the mentored stage of this award, I will begin to directly test my working hypothesis by implementing experimental and analytic techniques to quantify and interpret nanometer-size organization on the intact cell surface (Aim1). 1 will also develop experimental methods to modulate lateral heterogeneity in both plasma membrane vesicles and intact cell plasma membranes (Aim 2). Through these studies, I will acquire the skills and perspective necessary to successfully apply my strong physical background to this biomedically relevant area of research. As an independent researcher, I will continue work on both aims, eventually deciphering the physical basis of membrane lateral organization (Aim1), and developing novel methodologies to probe the functional consequences of lipid-mediated lateral heterogeneity in IgE mediated signaling in RBL mast cells (Aim 2). My long term goal is develop general methods to probe the functional roles of lipids in a wide range of processes and cell types which will enable new lines of study into the biomedical consequences of lipid mediated plasma membrane organization in cells. The Pathway to Independence Award will give me the resources to focus deeply on my biological training in my postdoctoral years, and the independence to pursue my own research objectives as I transition into my future position as a young faculty member. Relevance: It is widely accepted that plasma membrane lateral heterogeniety plays important functional roles in normal cell processes and in the development and maintenance of human diseases. I will open new opportunities for intervention in cell membrane-based pathologies by developing effective experimental techniques and a clear conceptual framework in which to study lipids and proteins in native membranes.