MAP BINDING EFFECT ON MICROTUBULE STIFFNESS

图谱结合对微管硬度的影响

• 批准号: 2459276
• 负责人: STEVEN P GROSS
• 金额: $ 2.86万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2459276
• 关键词: biophysics; chemical binding; microtubule associated protein; microtubules

The goal of this proposal is to explore further the interaction between microtubules (MTs) and microtubule-associated proteins (MAPs), particularly the ability of certain MAPs to alter the mechanical properties of MTs. It is possible that one of the key functions of MAP binding is to produce concomitant changes in MT stiffness, affecting neuronal development (MAP2 and Tau) or mitotic processes (MAP4), and there is preliminary evidence that MAP binding does result in an increase of MT stiffness. Details of how MAP binding changes MT mechanical properties, and how MAP phosphorylation alters such effects, are potentially useful in understanding certain neuronal diseases like Alzheimer's disease (where infected patients exhibit a hyperphosphorylated form of Tau), and may also help understand certain details of mitosis, during which MAP4's state of phosphorylation is changed by MPF. To accurately measure MT stiffness, a state-of-the-art dual-beam optical trap will be built for these experiments. The trap will be used to bend individual MTs, by exerting force on beads bound to each end of the MT. Included in the trap design is the ability to accurately measure applied force. Knowing the applied force and resultant curve in the MT will enable the determination of the MT stiffness. These measurements will be carried out in the absence and presence of MAPs, so that their effect on stiffness can be measured. Phosphorylated and genetically engineered MAPs will also be used in these studies, to examine their relative effects.

这项提议的目标是进一步探讨 微管(MT)和微管相关蛋白(MAP)， 尤其是某些地图改变机械结构的能力 MTS的属性。MAP的主要功能之一可能是 捆绑是产生随之而来的MT刚性变化，影响 神经元发育(MAP2和Tau)或有丝分裂过程(MAP4)，还有 是否有初步证据表明MAP结合确实会导致MT增加 僵硬。MAP绑定如何改变MT机械特性的详细信息， 以及MAP磷酸化如何改变这种效应，可能对 了解某些神经性疾病，如阿尔茨海默病(在哪里 感染的患者表现出一种过度磷酸化的Tau)，还可能 帮助了解有丝分裂的某些细节，在此期间，MAP4的S处于 MPF改变了磷酸化。为了准确地测量MT刚度，一个 最先进的双光束光学陷阱将为这些 实验。陷阱将被用来弯曲单个MT，通过施加 用力将珠子绑在MT的两端。包括在陷阱设计中 是精确测量外力的能力。了解适用对象 MT中的力和合成曲线将能够确定 MT刚度。这些测量将在不存在的情况下进行 贴图的存在，以便可以测量它们对刚度的影响。 磷酸化和基因工程地图也将用于这些 研究，以检查它们的相对影响。