STRUCTURAL STUDIES ON ANTIFREEZE PROTEINS

抗冻蛋白的结构研究

• 批准号: 6386661
• 负责人: FRANK D SONNICHSEN
• 金额: $ 11.92万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6386661
• 关键词: Coleoptera; Osteichthyes; cryoprotective agents; cryoscience; freezing; intermolecular interaction; nuclear magnetic resonance spectroscopy; protein structure function; proteins; site directed mutagenesis; water

Subfreezing temperatures pose a severe threat to the survival of organisms inhabiting polar and subpolar environments. Many organisms, including plants, insects and fish, have adapted and developed protective mechanisms which lower the risk of freezing. Certain fish produce antifreeze proteins, i.e. substances which lower the freezing points of their body fluids by binding to and inhibiting the growth of seed ice crystals. Similar compounds (thermal hysteresis proteins) are currently being isolated from plants and insects. Also, protein- crystal interactions are biologically common and important in processes such as biomineralization and calcification. This research project plans to investigate the structure-function relationship of several antifreeze proteins, characterize their unique activity and identify structural features or common motifs required for activity. The long- term objectives are to improve our understanding of the kinetic and thermodynamic aspects of the protein adsorption to ice in order to identify the source for the observed high binding affinity and mechanism of freezing point depression. The studies will increase our basic understanding on protein-surface and protein-crystal interactions, and are aimed to facilitate the engineering of antifreeze compounds for use in the cryogenic storage of cells, blood and organs, and for conferring freezing resistance to crop and food. My laboratory is dedicated to the structural and mechanistic characterization of a number of distinct classes of antifreeze proteins. The specific aims of this stage of the project will be to utilize homonuclear and heteronuclear multi-dimensional NMR-spectroscopy in combination with site-specific mutagenesis, computational methods and crystallization experiments to: i) determine the three dimensional structure of Type 2 antifreeze protein from herring, ii) determine the solution structure of Type 2 antifreeze protein from sea raven, iii) to determine the structure of a thermal hysteresis protein from the beetle Tenebrio molitor, and iv) to investigate the protein-water and protein-ice interactions of these proteins.

低于冰点的温度对人类的生存构成了严重威胁。 生活在极地和亚极地环境中的生物。 许多生物， 包括植物、昆虫和鱼类， 降低冷冻风险的保护机制。某些鱼类 产生抗冻蛋白质，即降低冻结的物质 通过结合并抑制其生长， 种下冰晶类似的化合物（热滞蛋白）是 目前从植物和昆虫中分离出来。 还有蛋白质 晶体相互作用在生物学上是常见的， 例如生物矿化和钙化。本研究项目 计划研究几个结构-功能关系 抗冻蛋白，表征其独特的活性，并确定 结构特征或活动所需的共同基序。很长的- 长期目标是提高我们对动力学的理解， 蛋白质吸附到冰的热力学方面， 确定观察到的高结合亲和力的来源， 凝固点降低的机理这些研究将增加我们的 对蛋白质表面和蛋白质晶体的基本认识 相互作用，并旨在促进防冻剂的工程 用于细胞、血液和器官的低温储存的化合物， 以及赋予农作物和食品抗冻性。我的实验室 是致力于结构和机械表征的一个 不同种类的抗冻蛋白。具体目标 该项目的这一阶段将是利用住房和 异向多维NMR光谱与 定点诱变、计算方法和结晶 i）确定型的三维结构 2.从鲱鱼中提取抗冻蛋白，ii）确定溶液结构 的2型抗冻蛋白，iii）以确定 拟步甲热滞蛋白的结构 molitor，以及iv）研究蛋白质-水和蛋白质-冰 这些蛋白质的相互作用。

项目成果

Alternative roles for putative ice-binding residues in type I antifreeze protein.

• DOI: 10.1021/bi982602p
• 发表时间: 1999-04
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: M. Loewen;H. Chao;M. Houston;J. Baardsnes;R. Hodges;C. Kay;B. Sykes;F. Sönnichsen;P. Davies

Characterization of the carboxylate delivery module of transcarboxylase: following spontaneous decarboxylation of the 1.3S-CO2- subunit by NMR and FTIR spectroscopies.

转羧酶的羧酸盐递送模块的表征：通过 NMR 和 FTIR 光谱对 1.3S-CO2- 亚基自发脱羧。

• DOI: 10.1021/bi0116442
• 发表时间: 2002
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Rivera-Hainaj,RosaE;Pusztai-Carey,Marianne;VenkatReddy,D;Choowongkomon,Kiattawee;Sönnichsen,FrankD;Carey,PaulR
• 通讯作者: Carey,PaulR