Novel Methodologies for the Preservation (and Recovery) of Proteins at Low Temperature

低温保存（和回收）蛋白质的新方法

• 批准号: 10088682
• 负责人: GUSTAVO E LOPEZ-QUINONES
• 金额: $ 11.91万
• 依托单位: HERBERT H. LEHMAN COLLEGE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-02 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10088682
• 关键词: Biological; Cell Death; Cells; Complex; Coupled; Cryopreservation; Environment; Exhibits; Food; Freezing; Genetic Polymorphism; Glass; Goals; Grant; Hemoglobin; Ice; Insulin; Knowledge; Lead; Link; Methodology; Methods; Modeling; Molecular; Pathway interactions; Peptides; Pharmacologic Substance; Phase; Procedures; Property; Protein Denaturation; Proteins; Recovery; Research; Solvents; Structure; System; Techniques; Temperature; Testing; Tissues; Ubiquitin; Water; Work; amorphous solid; aqueous; base; beta pleated sheet; biomaterial compatibility; cold temperature; crystallinity; density; design; experimental study; molecular dynamics; novel; preservation; pressure; prevent; protein complex; simulation

ABSTRACT The goal of this SC3 grant is to develop a novel method for the cryopreservation of proteins without the use of co-solvents, which are usually employed to limit the structural changes that can occur during freezing. The method exploits the glass polymorphism of water, i.e., low density amorphous (LDA) and high density amorphous (HDA) glasses can be produced following specific cooling/compression pathways. The pathways for storage and recovery of proteins will be studied using non-equilibrium molecular dynamic simulations coupled to all-atom force fields. The central hypothesis is that the density of the HDA will limit fluctuations in the solution upon cooling and the protein will not undergo any significant structural changes when vitrified in HDA. The rationale for the proposed research is that an understanding of the detailed mechanism by which proteins are preserved in glassy water, can lead to the rational design of experiments that allow for more controlled subzero protein preservation. In order to systematically develop this understanding three groups of systems, with increasing order of structural complexity will be studied: (i) short peptides with different secondary structures (ii) simple proteins that contain these secondary structures within their tertiary structures, and (iii) two biomedically relevant proteins. The information obtained from these studies has the potential to be extended to the storage of pharmaceuticals, food, and biological cells without the use of co-solvents.

摘要 SC 3资助的目标是开发一种蛋白质冷冻保存的新方法 而不使用共溶剂，共溶剂通常用于限制结构变化， 发生在冷冻期间。该方法利用了水的玻璃多晶型，即，低密度 非晶（LDA）和高密度非晶（HDA）玻璃可以按照特定的工艺制备， 冷却/压缩路径。研究蛋白质的储存和回收途径 使用非平衡分子动力学模拟耦合到所有原子力场。中央 假设HDA的密度将限制冷却时溶液中的波动， 当在HDA中玻璃化时，蛋白质不会经历任何显著的结构变化。的理由 对于拟议的研究是，了解蛋白质的详细机制， 保存在玻璃水中，可以导致合理的实验设计，允许更多的 受控的低温蛋白质保存。为了系统地发展这种理解 三组系统，随着结构复杂性的增加，将被研究：（一）短 具有不同二级结构的肽（ii）含有这些二级结构的简单蛋白质 它们的三级结构内的结构，和（iii）两个生物医学相关的蛋白质。的 从这些研究中获得的信息有可能扩展到存储 在不使用共溶剂的情况下用于药物、食品和生物细胞。