Single molecule tracking of qdot-conjugated SNAREs in planar supported bilayers

平面支撑双层中 qdot 共轭 SNARE 的单分子追踪

• 批准号: 7222278
• 负责人: Alfred S Kwok
• 金额: $ 3.76万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-12-01 至 2007-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7222278
• 关键词: Address; Automobile Driving; Behavior; Cell membrane; Chimeric Proteins; Complex; Condition; Data; Development; Diffusion; Disease; Docking; Dopamine; Drug Delivery Systems; Exhibits; Experimental Designs; Functional disorder; Goals; Hormones; Individual; Lateral; Lead; Lipids; Liquid substance; Maleimides; Melanocytic nevus; Membrane; Membrane Fusion; Membrane Microdomains; Mental Depression; Mole the mammal; Motion; Neurons; Neurotransmitters; PC12 Cells; Pharmacotherapy; Phase; Phospholipids; Property; Proteins; Quantum Dots; Range; Research; Role; SNAP receptor; Signal Transduction; Transport Vesicles; Vesicle; addiction; design; neuropsychiatry; particle; preference; presynaptic; research study; single molecule; soluble NSF attachment protein; target SNARE proteins

DESCRIPTION (provided by applicant): The long range goal for this research is to perform single particle tracking with quantum dots (qdots) to further our understanding of the role of SNARE proteins in neuronal membrane fusion. Presynaptic fusion is a key step in vesicle transport, and vesicle transport is involved in the delivery of vital neurotransmitters such as dopamine and other secretory hormones that are important in signaling and development. Further understanding of presynaptic fusion will enable better understanding of vesicle transport and thus the pathophysiology of neuropsychiatric conditions such as depression and addiction. In addition, understanding the functioning of SNAREs may even lead to the design of better drugs/therapy as one envisions enlisting SNAREs for fusion-enhanced targeted drug delivery Single particle tracking (SPT) of qdots will be performed to study the lateral diffusion of t-SNAREs on supported bilayer membranes with raft-forming lipids to address the inconsistent observations involving the phase preference for t-SNAREs in several recent studies. Proof-of-principle experiments will first be performed to optimize the experimental design and setup and to ascertain the efficacy of using qdots for single particle tracking. SPT of qdot-conjugated lipid will be used to study lipid diffusion in supported homogenous bilayers and heterogeneous bilayers with raft-forming lipids. t-SNARE diffusion in supported heterogenous bilayers will then be studied using SPT of qdot-conjugated SNAREs. Different mole fractions of t-SNAREs and different raft-forming lipid mixtures will be used, and cumulative distribution function will be used to fit the data to reveal multi-component diffusive behavior.

描述（由申请人提供）：本研究的长期目标是使用量子点（qdots）进行单粒子跟踪，以进一步了解SNARE蛋白在神经元膜融合中的作用。突触前融合是囊泡运输的关键步骤，而囊泡运输涉及重要神经递质的传递，如多巴胺和其他在信号传导和发育中重要的分泌激素。进一步了解突触前融合将有助于更好地理解囊泡运输，从而更好地理解抑郁和成瘾等神经精神疾病的病理生理学。此外，了解SNAREs的功能甚至可能导致设计更好的药物/治疗，因为有人设想将SNAREs用于融合增强靶向药物递送，将进行量子点的单粒子跟踪（SPT）来研究t-SNAREs在具有筏状形成脂质的支持双层膜上的横向扩散，以解决最近几项研究中关于t-SNAREs相偏好的不一致观察。首先将进行原理验证实验，以优化实验设计和设置，并确定使用量子点进行单粒子跟踪的有效性。量子点共轭脂质的SPT将用于研究脂质在支持的均匀双分子层和具有筏状脂质的非均匀双分子层中的扩散。t-SNARE在支持的异质双层中的扩散将使用量子点共轭SNAREs的SPT进行研究。将使用不同的t-SNAREs摩尔分数和不同的筏形脂质混合物，并使用累积分布函数对数据进行拟合，以揭示多组分扩散行为。