A new tool for measuring surface-biomolecule interactions

测量表面生物分子相互作用的新工具

基本信息

批准号：
8823777
负责人：
Kevin W Plaxco
金额：
$ 22.33万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA BARBARA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8823777
关键词：
Acute Address Adsorption Amines Amino Acids Behavior Binding Binding Proteins Biocompatible Materials Biological Biological Phenomena Biopolymers Biosensor Biotechnology Cell membrane Charge Chimera organism DNA Development Devices Drug Targeting Electrodes Empirical Research Exhibits Free Energy Gold Health Investigation Ionic Strengths Length Literature Maps Measurement Measures Membrane Proteins Methods Methylene blue Modeling Monitor Nature Nucleic Acid Binding Nucleic Acids Optical reporter Outcome Oxidation-Reduction Peptides Phase Physiologic pulse Protein Microchips Proteins Reporter Research Resistance Site Solutions Structure Surface Techniques Tertiary Protein Structure Testing Thermodynamics Thymine Time Tissue Engineering Urea Validation Variant Work base density design improved inorganic phosphate insight interest migration monolayer nanoparticle novel phosphonate polypeptide predictive modeling programs research study response scaffold stem theories tool

项目摘要

DESCRIPTION (provided by applicant): Why do biomolecules retain activity when attached to some surfaces (e.g., the cell membrane) and yet almost invariably unfold and inactivate when attached to others (e.g., many artificial surfaces)? And how can we recreate the former effect to produce artificial surfaces on which attached biomolecules similarly retain their function? To date our ability to address these important questions has been hampered by an acute lack of quantitative experimental methods for measuring the thermodynamics of biomolecule-surface interactions. That is, despite a large body of qualitative literature describing how adsorption alters biomolecular structure, and a large number of empirical studies searching for adsorption- resistant surfaces, quantitative, experimentally testable insights into how and why this occurs have proven elusive. Thus motivated, we propose here the development and validation of a novel experimental (electrochemical) approach to measuring the folding free energy of biomolecules site-specifically attached to well-defined macroscopic surfaces. Comparison with the folding free energy in bulk solution then informs on the thermodynamics -and thus mechanisms- underlying biopolymer-surface interactions. To date we have employed this approach to characterize the easily modeled folding of a surface-confined DNA stem-loop in studies that have, for the first time, defined experimentally the extent to which, and mechanisms by which a specific biomolecule interacts with a range of well-defined, macroscopic surfaces. Here we propose a two-year research program aimed at adapting this quantitative experimental tool to the study of protein-surface interactions.

描述（由申请人提供）：当附着在某些表面（例如细胞膜）上时，生物分子为什么会保留活性，而当与其他表面相连（例如，许多人造表面）时几乎总是会展开并灭活？我们如何重新创建以前的效果以产生人工表面，其中附着的生物分子类似地保留其功能？迄今为止，我们解决这些重要问题的能力受到急性缺乏定量实验方法来测量生物分子 - 表面相互作用的热力学。也就是说，尽管有大量的定性文献描述了吸附如何改变生物分子结构，以及大量的经验研究寻找吸附性 - 耐药表面，但定量，可以实验测试的见解，即如何以及为什么出现这种情况证明这是错误的。因此，我们在这里提出了一种新型实验（电化学）方法的开发和验证，以测量特定于定义明确定义的宏观表面的生物分子位点的折叠自由能。与散装溶液中的折叠自由能进行比较，然后告知热力学，从而提供了基础生物聚合物 - 表面相互作用的机制。迄今为止，我们已经采用了这种方法来表征表面限制的DNA茎环的易于建模的研究，这些研究首次在实验上定义了特定生物分子与一系列良好定义的宏观表面相互作用的机制。在这里，我们提出了一项为期两年的研究计划，旨在将此定量的实验工具调整为蛋白质表面相互作用的研究。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Experimental Measurement of Surface Charge Effects on the Stability of a Surface-Bound Biopolymer.

表面电荷对表面结合生物聚合物稳定性影响的实验测量。

DOI：
10.1021/acs.langmuir.8b01004
发表时间：
2018
期刊：
Langmuir : the ACS journal of surfaces and colloids
影响因子：
0
作者：
Watkins,HerschelM;Ricci,Francesco;Plaxco,KevinW
通讯作者：
Plaxco,KevinW

Quantitative measurements of protein-surface interaction thermodynamics.

蛋白质-表面相互作用热力学的定量测量。

DOI：
10.1073/pnas.1800287115
发表时间：
2018
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Kurnik,Martin;Ortega,Gabriel;Dauphin-Ducharme,Philippe;Li,Hui;Caceres,Amanda;Plaxco,KevinW
通讯作者：
Plaxco,KevinW

Effects of crowding on the stability of a surface-tethered biopolymer: an experimental study of folding in a highly crowded regime.