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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：
8662567
负责人：
Kevin W Plaxco
金额：
$ 18.58万
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 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8662567
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 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 public health relevance research study response scaffold stem theories tool

项目摘要

Project summary. 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.

项目总结。为什么生物分子附着在某些表面（如细胞）时仍能保持活性