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）：为什么生物分子在附着到某些表面（例如细胞膜）时保留活性，而在附着到其他表面（例如许多人造表面）时几乎总是展开并失活？我们如何才能重现前一种效果，以产生附着的生物分子同样保留其功能的人造表面？迄今为止，由于严重缺乏测量生物分子-表面相互作用的热力学的定量实验方法，我们解决这些重要问题的能力受到阻碍。也就是说，尽管大量定性文献描述了吸附如何改变生物分子结构，并且进行了大量寻找抗吸附表面的实证研究，但对于这种现象如何发生以及为何发生的定量、可通过实验测试的见解已被证明是难以捉摸的。因此，我们在此提出开发和验证一种新颖的实验（电化学）方法来测量特定位点附着在明确的宏观表面上的生物分子的折叠自由能。然后与本体溶液中的折叠自由能进行比较，了解热力学以及生物聚合物表面相互作用的机制。迄今为止，我们已经在研究中采用这种方法来描述表面限制 DNA 茎环的容易建模的折叠，这些研究首次通过实验定义了特定生物分子与一系列相互作用的程度和机制。定义明确的宏观表面。在这里，我们提出了一个为期两年的研究计划，旨在使这种定量实验工具适应蛋白质-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.