Net charge in protein structure and aggregation: a charge ladder approach

蛋白质结构和聚集中的净电荷：电荷梯方法

• 批准号: 7275003
• 负责人: Bryan F. Shaw
• 金额: $ 4.68万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7275003
• 关键词: Alzheimer' s Disease; Amides; Anions; Biophysics; Charge; Chemicals; Disease; Electrostatics; Future; Hydrogen; Hydroxide Ion; Hydroxides; Investigation; Link; Malignant Neoplasms; Mass Spectrum Analysis; Membrane Lipids; Membrane Proteins; Modification; Molecular; Nanotechnology; Neurodegenerative Disorders; Non-Insulin-Dependent Diabetes Mellitus; Pathogenesis; Pattern; Pharmacotherapy; Play; Post-Translational Protein Processing; Printing; Property; Proteins; Public Health; Range; Rate; Reaction; Research; Role; Sorting - Cell Movement; Spectrometry, Mass, Electrospray Ionization; Structure; Surface; Technology; Testing; concept; human disease; monolayer; novel strategies; prevent; protein aggregation; protein folding; protein misfolding; protein structure; research study; therapy development; tool

DESCRIPTION (provided by applicant): This proposal describes ongoing and future-planned investigations that seek to determine the role that net charge plays in biophysics-related problems. The first aim presents the hypothesis that a protein's net charge plays a significant role in determining the rate of amide hydrogen exchange in proteins - a reaction that is catalyzed by hydroxide anion at pH>4. Protein charge ladders and mass spectrometry are used to test this charge-dependent exchange hypothesis. The second aim of this proposal hypothesizes that protein aggregation, as related to many human diseases, can be inhibited with subtle chemical modifications that increase the protein's net charge. Controlling charge - as opposed to some other parameter or property such as native-state stability or structure - is an underemphasized and unexplored possibility in developing drug therapies for a wide array of diseases linked to protein aggregation. The experiments proposed in Aim 2 are intended as a proof of concept for this approach (termed PRO-CHAIN; PROtein CHarging for Aggregation INhibition). Aim 3 proposes experiments to explore if electrostatic interactions between proteins and surfaces can be used to i) promote ii) direct iii) pattern, or iv) inhibit protein aggregation onto charge- micro-patterned self assembled monolayers (SAMs). The aggregation of proteins onto charged surfaces, such as lipid membranes, is gaining acceptance as a likely scenario in disease pathogenesis. This third aim describes experiments using protein charge ladders, micro-contact printing and self assembled monolayers to explore this important phenomenon that is suspected to be relevant to a wide range of diseases. Relevance to public health: This proposal has immediate relevance to public health; first, it describes a whole new approach for developing drug therapies, referred to as PRO-CHAIN (PROtein CHarging for Aggregation INhibition), that may aid in the development therapies for the treatment of a wide range of protein aggregation diseases such as Alzheimer's disease and type II diabetes. Furthermore, this proposal describes research that may further our understanding of the molecular determinants of hydrogen exchange in protein folding studies -- many of these studies focus on pathogenic proteins that cause human disease. Lastly, this proposal uses new cutting edge technologies to study protein-surface interactions of the sort that may be related to numerous neurodegenerative diseases, as well as cancer.

描述（由申请人提供）：该提案描述了正在进行的和未来计划的调查，这些调查旨在确定净电荷在与生物物理相关问题中的作用。第一个目的提出了以下假设：蛋白质的净电荷在确定蛋白质中酰胺氢交换的速率中起重要作用 - 这种反应是在pH> 4时由氢氧化阴离子催化的反应。蛋白质电荷梯子和质谱法用于检验该电荷依赖性交换假设。该提案的第二个目的假设，与许多人类疾病有关的蛋白质聚集可以通过细微的化学修饰来抑制，从而增加蛋白质的净电荷。控制电荷 - 与其他参数或财产（例如天然状态稳定性或结构）相反，在开发与蛋白质聚集有关的多种疾病的药物疗法方面，是一种未经探索和未开发的可能性。 AIM 2中提出的实验旨在作为这种方法的概念证明（称为亲链；用于聚集抑制的蛋白质充电）。 AIM 3提出了实验，以探索蛋白质和表面之间的静电相互作用是否可以用于i）促进ii）ii）直接iii）模式，或iv）抑制蛋白质聚集到电荷 - 微图案的自组装单层（SAM）。蛋白质在带电表面（例如脂质膜）上的聚集正在接受，这可能是疾病发病机理的一种情况。第三个目标描述了使用蛋白质电荷梯子，微接触印刷和自组装单层的实验，以探索这种重要现象，该现象被认为与多种疾病有关。与公共卫生的相关性：该提案与公共卫生有直接的相关性；首先，它描述了一种用于开发药物疗法的全新方法，该方法称为亲链（蛋白质抑制蛋白质抑制），可能有助于开发疗法，以治疗多种蛋白质聚集疾病，例如阿尔茨海默氏病和II型糖尿病。此外，该提案描述了可能进一步了解蛋白质折叠研究中氢交换的分子决定因素的研究 - 其中许多研究集中于引起人类疾病的致病蛋白。最后，该提案使用新的尖端技术来研究可能与许多神经退行性疾病以及癌症有关的蛋白质表面相互作用。