A Novel Technique of Imposing Femtonewton Forces

一种施加飞牛顿力的新技术

• 批准号: 7036067
• 负责人: JIN-YU SHAO
• 金额: $ 28.65万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7036067
• 关键词: biophysics; cell adhesion; cell line; computer human interaction; computer simulation; computer system design /evaluation; intermolecular interaction; ligands; mathematics; molecular dynamics; nanotechnology; protein folding; protein protein interaction; protein structure function; selectins; technology /technique development; video microscopy

DESCRIPTION (provided by applicant): Protein deformation, protein folding, and protein interaction with other proteins (e.g., receptor-ligand binding) are all mediated by piconewton or femtonewton level forces. Only one of the existing low force application techniques can be used to impose forces of femtonewton level. Therefore, the overall goal of this proposal is to develop and validate a novel technique of imposing femtonewton level forces. The goal of the R21 phase of this proposal is to conduct a comprehensive numerical analysis and an experimental validation of the proposed technique (EMAT) to assess its feasibility. The specific aims of the R21 phase are: 1) to conduct an axisymmetric analysis of the EMAT, 2) to conduct a three-dimensional analysis of the EMAT, and 3) to validate the EMAT with ultra-soft springs. The goal of the R33 phase of this proposal is to build a database that can be used by other researchers to calculate forces for various combinations of all the parameters involved, to build a device that can be used to further validate the EMAT experimentally, and to apply it to single receptor-ligand interactions. The specific aims of the R33 Phase are: 1) to develop a computer program that can be used by others to easily calculate the force imposed with the EMAT, 2) to implement a computer-controlled micropipette manipulation system that can be used to impose femtonewton level forces, 3) to further validate the EMAT with optical tweezers, and 4) to examine the effect of force on the interaction between L-selectin and its ligands. The analyses proposed in the R21 phase will be performed with FIDAP (a finite element analysis software package for computational fluid dynamics) to evaluate the effects of some critical parameters and the alignment between the micropipette and transducer on the calculated force. The validation of the EMAT will be carried out with the micropipette manipulation system, ultra-soft springs, and optical tweezers. Combined with the micropipette aspiration technique, the EMAT will enable us to impose a wide range of forces from a few femtonewtons to about 100 nanonewtons, which is unprecedented. Therefore, the EMAT should have great potential to make an impact in the field of single molecule and single bond biophysics.

描述（由申请人提供）： 蛋白质变形，蛋白质折叠和与其他蛋白质的相互作用（例如受体 - 配体结合）均由piconewton或femtonewton水平力介导。 现有的低力应用技术中只能使用一种施加femtonewton级别的力。 因此，该提案的总体目标是开发和验证一种新型的施加femtonewton水平力量的技术。 该提案的R21阶段的目的是进行全面的数值分析和对拟议技术（EMAT）的实验验证，以评估其可行性。 R21相的具体目的是：1）对EMAT进行轴对称分析，2）对EMAT进行三维分析，以及3）用超柔软的弹簧验证EMAT。 该提案的R33阶段的目的是构建一个数据库，该数据库可以被其他研究人员用于计算所有涉及所有参数的各种组合的力，以构建可用于进一步验证EMAT实验验证的设备，并将其应用于单个受体 - 配体相互作用。 R33阶段的具体目的是：1）开发其他人可以用来轻松计算emat施加的力的计算机程序，2）实施计算机控制的微孔操纵系统，该系统可用于强加于fomtonewton级别的力量，3）用光学的镊子进一步验证emat，并检查效应的效果，并效应效果。 R21阶段提出的分析将使用FIDAP（用于计算流体动力学的有限元分析软件包）来评估某些关键参数的效果以及微孔和传感器对计算力的效果。 EMAT的验证将使用微型移动系统，超柔软的弹簧和光学镊子进行。结合微管抽吸技术，EMAT将使我们能够从几个femtonewtons到大约100个纳米温顿，这是前所未有的。 因此，EMAT应该具有在单分子和单键生物物理学领域产生影响的巨大潜力。