Structural-functional characterization of a hyperactive antifreeze protein

高活性抗冻蛋白的结构功能表征

• 批准号: 1413696
• 负责人: Krisztina Varga
• 金额: $ 55万
• 依托单位: University of Wyoming
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1413696&HistoricalAwards=false
• 关键词: Structural; functional; characterization; hyperactive; antifreeze

With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding the research of Dr. Krisztina Varga and Dr. Daniel Levy from the University of Wyoming to structurally and functionally characterize a hyperactive insect antifreeze protein. Antifreeze proteins are produced in several organisms that live in cold climates and contribute to their resistance to freezing. They bind irreversibly to the surface of ice crystals and inhibit their growth, however, the exact mechanism at the molecular level is not fully understood. The investigators will determine the structure of the antifreeze protein, examine its water binding properties, and investigate its mechanism of freeze inhibition in live cells. The results of this research will provide a better fundamental understanding of molecular processes characteristic of antifreeze proteins interacting with ice and expand their practical applications, for example, in protecting foods from damage by freezing. The project will have a broader impact by providing opportunities for underrepresented groups of graduate and undergraduate students to participate in scientific research, many of them being first generation university students from rural areas. The project will be integrated into outreach programs for community college, high school and grade school students.This research will probe the structure and mechanism of action of the hyperactive insect antifreeze protein ApAFP752. The research will provide a better understanding of how antifreeze proteins interact with ice and provide cryoprotection to organisms that live in cold environments. The approaches are comprehensive and combine biochemistry, NMR spectroscopy, and confocal microscopy for both in vitro and in vivo studies and will utilize the synergy of the expertise of the investigators in these areas. This research project will be undertaken to (1) structurally characterize the antifreeze protein by NMR spectroscopy, (2) explore its water binding properties and interactions (e.g. hydrogen bonding), and (3) investigate its cryoprotective mechanism in Xenopus live cells. The comprehensive approach will provide unique insights into the underlying molecular mechanism of cryopreservation.This project is co-funded by the Experimental Program for the Stimulation of Competitive Research (EPSCoR).

有了这个奖项，化学部的生命过程化学项目资助了怀俄明州大学的Krisztina Varga博士和丹尼尔利维博士的研究，以从结构和功能上表征一种过度活跃的昆虫抗冻蛋白。抗冻蛋白在生活在寒冷气候中的几种生物体中产生，有助于它们抵抗冷冻。它们与冰晶表面不可逆地结合并抑制其生长，然而，在分子水平上的确切机制尚未完全了解。研究人员将确定抗冻蛋白的结构，检查其水结合特性，并研究其在活细胞中的冷冻抑制机制。这项研究的结果将提供对防冻蛋白与冰相互作用的分子过程特征的更好的基本了解，并扩大其实际应用，例如，在保护食品免受冷冻损坏方面。该项目将产生更广泛的影响，为代表性不足的研究生和本科生群体提供参与科学研究的机会，其中许多人是来自农村地区的第一代大学生。该项目将被整合到社区大学，高中和小学学生的推广计划中。本研究将探索高度活跃的昆虫抗冻蛋白ApAFP752的结构和作用机制。这项研究将更好地了解抗冻蛋白如何与冰相互作用，并为生活在寒冷环境中的生物提供低温保护。这些方法是全面的，并结合了联合收割机生物化学，核磁共振光谱学和共聚焦显微镜在体外和体内研究，并将利用这些领域的研究人员的专业知识的协同作用。该研究项目将进行（1）通过NMR光谱法表征抗冻蛋白的结构，（2）探索其水结合特性和相互作用（例如氢键），以及（3）研究其在爪蟾活细胞中的冷冻保护机制。综合的方法将提供独特的见解的潜在的分子机制的冷冻保存。该项目是共同资助的实验计划刺激的竞争性研究（EPSCoR）。