IDENTIFICATION AND MOLECULAR BASIS FOR EFFICIENT ANTIFREEZE PROTIEN ENHANCERS

高效抗冻蛋白增强剂的鉴定和分子基础

• 批准号: 8266184
• 负责人: Xin Wen
• 金额: $ 10.88万
• 依托单位: CALIFORNIA STATE UNIVERSITY LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8266184
• 关键词: Adsorption; Affect; Affinity; Amines; Amino Acids; Antifreeze; Antifreeze Proteins; Binding; Biological Assay; Chemicals; Circular Dichroism Spectroscopy; Conflict (Psychology); Cryopreservation; Cryosurgery; Data; Depressed mood; Development; Enhancers; Exhibits; Fish Type I APF; Fishes; Food Preservation; Freezing; Frozen Foods; Funding; Goals; Health; Heterocyclic Compounds; Human; Hydrogen; Hydrogen Bonding; Hydrophobic Interactions; Ice; Insecta; Knowledge; Measures; Methods; Modification; Molecular; NMR Spectroscopy; Nature; Nitrogen; Order Coleoptera; Organism; Plants; Play; Property; Public Health; Reporting; Research; Role; Series; Site-Directed Mutagenesis; Solutions; Structure; Surface; System; Temperature; Tenebrio; Time; Tissue Preservation; Variant; Water; Work; base; biological systems; cold temperature; electron donor; improved; insight; macromolecule; melting; molecular mass; practical application; solute

DESCRIPTION (provided by applicant): Antifreeze proteins (AFPs) have been identified in a number of organisms, such as fish, plants, and insects, to allow them to survive at subfreezing temperatures. They are all characterized by their ability to depress the freezing point of the solution without appreciably altering the melting point thereby producing a thermal hysteresis. Insect AFPs are often the most active AFPs (10-100 times more active than type I fish AFPs). The antifreeze activities of many AFPs can be further enhanced in the presence of certain co-solutes (called enhancers). Both low molecular mass chemicals and macromolecules have been identified as enhancers for AFPs and some of the enhancers do play a role in enhancing the antifreeze activity of AFPs physiologically. Some of the enhancers have been reported to bind AFPs specifically. The mechanism of the enhancement effect, however, is still poorly described. The AFP from the beetle Dendroides canadensis (DAFP) and the enhancers have been extensively studied. The binding of enhancers to AFP has been demonstrated as one important mechanism for the enhancement effect of enhancers on the antifreeze activity of AFPs. Is there a common mechanism for the enhancement phenomena in different AFP systems? In this project, we will examine an important beetle AFP system, a mealworm AFP from Tenebrio molitor (TmAFP) and the enhancers. We propose that the enhancement efficiency of the enhancers for TmAFP antifreeze activity depends on the physiochemical properties of the enhancers. We will assess the enhancement efficiency of selected amine derivatives on TmAFP antifreeze activity and correlate their enhancement efficiency with their differing physicochemical properties. We will identify the possible interactions between TmAFP and the enhancers and determine the key amino acids in TmAFP involving in binding to the enhancers. The study of AFPs and enhancers is essential to advance our understanding of the biological system and facilitate the development of highly efficient AFP systems for their practical biomedical applications, such as in preservation of tissues at low temperatures in the lab and in human health as well as other industrial applications, such as in frozen foods and in hydrate inhibition. The proposed research will provide insights into the molecular mechanism of AFP and AFP enhancers and resolve conflicting reports regarding the mechanism of action of AFPs. PUBLIC HEALTH RELEVANCE: The proposed work examines the role of "enhancers" on the antifreeze properties of antifreeze proteins (AFPs). Antifreeze protection is important in public health (e.g., preservation of tissues at low temperatures and food preservation). The results of the project will advance our understanding of how AFPs and enhancer work, and improve public health by facilitating the biomedical applications of AFPs and enhancers.

描述（由申请人提供）：抗冻蛋白（AFP）已在许多生物体（如鱼类、植物和昆虫）中鉴定，使其能够在冰点以下的温度下存活。它们都具有降低溶液凝固点而不明显改变熔点从而产生热滞后的能力。昆虫AFP通常是最活跃的AFP（比I型鱼类AFP活跃10-100倍）。许多AFP的抗冻活性可以在某些共溶质（称为增强剂）的存在下进一步增强。低分子量化合物和大分子物质都被认为是抗冻蛋白的增强剂，其中一些增强剂在生理上确实起到增强抗冻蛋白活性的作用。一些增强子已被报道特异性结合AFP。然而，增强效应的机制仍然很难描述。加拿大石斛甲胎蛋白（Dendroides canadensis，DAFP）及其增强剂已被广泛研究。增强子与AFP的结合已被证明是增强子增强AFP抗冻活性的重要机制之一。不同AFP系统中的增强现象是否存在共同的机制？本研究以黄粉虫甲胎蛋白（TmAFP）为研究对象，对一种重要的甲虫甲胎蛋白系统及其增强子进行了研究。我们认为，增强剂对TmAFP抗冻活性的增强效率取决于增强剂的理化性质。我们将评估所选胺衍生物对TmAFP抗冻活性的增强效率，并将其增强效率与其不同的物理化学性质相关联。我们将鉴定TmAFP与增强子之间可能的相互作用，并确定TmAFP中参与与增强子结合的关键氨基酸。AFP和增强剂的研究对于促进我们对生物系统的理解和促进高效AFP系统的开发对于其实际生物医学应用至关重要，例如在实验室和人类健康中低温保存组织以及其他工业应用，例如冷冻食品和水合物抑制。这项拟议的研究将为AFP和AFP增强剂的分子机制提供见解，并解决有关AFP作用机制的相互矛盾的报道。 公共卫生相关性：拟议的工作审查的作用“增强剂”的抗冻蛋白（AFP）的抗冻性能。防冻保护在公共卫生中很重要（例如，低温下的组织保存和食物保存）。该项目的结果将促进我们对AFP和增强剂如何工作的理解，并通过促进AFP和增强剂的生物医学应用来改善公众健康。