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

基本信息

批准号：
10460399
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
项目类别：
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/10460399
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 experimental study molecular dynamics novel preservation pressure prevent protein complex rational design simulation

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 experimental study molecular dynamics novel preservation pressure prevent protein complex rational design simulation

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项目摘要

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.

抽象的 SC3赠款的目的是开发一种新的蛋白质冷冻保存方法没有使用共渗透的情况，通常使用这些溶剂来限制可以限制的结构变化在冻结期间发生。该方法利用水的玻璃多态性，即低密度可以在特定之后产生无定形（LDA）和高密度无定形（HDA）眼镜冷却/压缩途径。将研究蛋白质的存储和恢复途径使用与全原子力场耦合的非平衡分子动力学模拟。中央假设是HDA的密度将在冷却后限制溶液中的波动当在HDA中玻璃化时，蛋白质不会发生任何显着的结构变化。理由对于拟议的研究，是对蛋白质的详细机制的理解保存在玻璃水中，可以导致实验的合理设计，从而允许更多控制的亚零蛋白保存。为了系统地发展这种理解将研究三组系统，结构复杂性的序列增加：（i）简短具有不同二级结构的肽（II）简单蛋白质，其中包含这些次级其三级结构内的结构，以及（iii）两个生物医学相关的蛋白质。这从这些研究中获得的信息有可能扩展到存储药物，食物和生物细胞无共溶液。