Experimental and Modeling Study of the Optical Properties and Phase Behavior of Heat and Cold Tolerant Eye Lens Crystallins

耐热冷晶状体晶状体光学性质和相行为的实验与模拟研究

• 批准号: 2003837
• 负责人: Rachel Martin
• 金额: $ 52.5万
• 依托单位: University of California-Irvine
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-15 至 2024-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2003837&HistoricalAwards=false
• 关键词: Experimental; Modeling; Study; Optical; Properties

Non-technical summaryThe objective of this project is to understand and mimic the properties of structural proteins that are adapted to resist extreme environments. A combination of experimental and theoretical techniques will be used to explore the molecular mechanisms by which eye lens proteins from different organisms maintain transparency. The scientific goal is to understand how nature produces transparent, durable biomaterials, as exemplified by the proteins making up the eye lens. The lenses of animals that live in very hot, cold, or UV light-saturated environments can provide a guide to making robust, biodegradable materials that withstand harsh conditions. The knowledge gained from this project will guide the design of flexible materials with potential applications to new biomimetic lens designs, in both the medical and analytical chemistry contexts, flexible microlenses, and self-healing materials that resist UV-light damage. In the future, it could enable the development of bioinorganic hydrogel scaffolds, controllable membraneless bioreactors, and ocean-safe, biodegradable sunscreen. The educational plan includes training of students at the Ph.D. and undergraduate levels. These students will be trained not only in the specific skills needed to complete the research, but also in critical thinking, scientific communication, and problem solving across disciplinary boundaries. Another important component is outreach to middle school and high school students in grades 6-12. The goal of the outreach program is to increase interest in science as a career among middle school students. The students, many of whom are from disadvantaged economic backgrounds, visit UCI to participate in a campus tour where they learn about undergraduate life, perform hands-on activities in the investigators’ laboratories, and interact with undergraduate researchers.  Technical summaryThe objective of this project is to understand the solubility and stability of proteins that are adapted to resist extreme environments. Experimental and modeling studies of highly stable structural proteins that play the same functional role in very different environments will provide insight into sequence determinants of stability and solubility. Structural and refractive proteins are chosen specifically to avoid interference from sequence features that are conserved to maintain chemical activity in thermostable enzymes. The scientific goal of this project is to understand how nature produces transparent, durable biomaterials, as exemplified by the proteins making up the eye lens. Although the molecular details of these proteins vary among different organisms, they are functionally similar in their high stability and resistance to temperature extremes. The specific proteins chosen for this effort are eye lens crystallins that are adapted to extreme environments. The J2 crystallin from the box jellyfish Tripedalia cystophora and the βγ-crystallin from the tunicate Ciona intestinalis are extremely stable with respect to heat denaturation. These proteins are not homologous but share a common function; we seek to understand the sequence and structural determinants of thermal stability. On the other hand, the γS- and γM-crystallins from the Antarctic toothfish (Dissostichus mawsoni) resist cold cataract at -2 C or lower, far below the temperature at which mammalian lenses undergo cold cataract. These proteins exist in a complex mixture, with 13 paralogs in the fish lens. One goal is to understand and control the liquid-liquid phase separation involved in cold cataract. The iota-crystallin from the diurnal gecko Lygodactylus picturatus binds a modified form of the visual pigment retinol to protect its retinae from UV light damage in bright sunlight. NMR and optical spectroscopy, light scattering, and directed mutagenesis experiments will be coupled with molecular dynamics and Monte Carlo simulations to elucidate the important structural factors and intermolecular interactions that confer high stability and resistance to extreme conditions on these proteins, guiding the future design of biomimetic materials. Understanding the structural factors diverse lens proteins have in common will help guide the design of future temperature- and UV-resistant biomimetic materials. Furthermore, the ability to predict and control the formation and dissociation of liquid-liquid phase separation in protein mixtures will be useful for modulating protein aggregation and biochemical activity.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

非技术摘要该项目的目标是了解和模拟适应极端环境的结构蛋白的特性。 将结合实验和理论技术来探索来自不同生物体的眼晶状体蛋白保持透明度的分子机制。 科学目标是了解大自然如何产生透明、耐用的生物材料，例如构成眼睛晶状体的蛋白质。生活在极热、极冷或紫外线饱和环境中的动物的晶状体可以为制造能够承受恶劣条件的坚固、可生物降解的材料提供指导。 从该项目中获得的知识将指导柔性材料的设计，这些材料可能应用于新的仿生透镜设计，包括医学和分析化学环境、柔性微透镜以及抵抗紫外线损伤的自修复材料。未来，它可以促进生物无机水凝胶支架、可控无膜生物反应器和海洋安全、可生物降解防晒霜的开发。教育计划包括博士生的培训。和本科水平。这些学生不仅将接受完成研究所需的特定技能的培训，还将接受批判性思维、科学沟通和跨学科边界解决问题的培训。 另一个重要组成部分是向 6 至 12 年级的初中和高中学生进行宣传。该推广计划的目标是提高中学生对科学作为职业的兴趣。这些学生中许多来自贫困的经济背景，他们访问加州大学尔湾分校参加校园参观，了解本科生生活，在研究人员的实验室进行实践活动，并与本科生研究人员互动。  技术摘要该项目的目标是了解适应极端环境的蛋白质的溶解度和稳定性。 对在非常不同的环境中发挥相同功能作用的高度稳定的结构蛋白进行实验和建模研究将有助于深入了解稳定性和溶解度的序列决定因素。专门选择结构蛋白和折射蛋白，以避免来自序列特征的干扰，这些序列特征是为了维持热稳定酶的化学活性而保守的。该项目的科学目标是了解大自然如何产生透明、耐用的生物材料，例如构成眼睛晶状体的蛋白质。尽管这些蛋白质的分子细节在不同生物体之间有所不同，但它们在功能上相似，具有高稳定性和对极端温度的抵抗力。为此目的选择的特定蛋白质是适应极端环境的晶状体蛋白。来自箱形水母 Tripedalia Cytophora 的 J2 晶状体蛋白和来自被囊动物 Ciona Enteris 的 βγ-晶状体蛋白对于热变性极其稳定。这些蛋白质不是同源的，但具有共同的功能；我们试图了解热稳定性的顺序和结构决定因素。另一方面，来自南极齿鱼（Dissostichus mawsoni）的γS-和γM-晶状体蛋白在-2℃或更低的温度下可以抵抗冷性白内障，远低于哺乳动物晶状体发生冷性白内障的温度。这些蛋白质以复杂的混合物形式存在，鱼晶状体中有 13 种旁系同源物。目标之一是了解和控制冷白内障涉及的液-液相分离。来自昼行性壁虎Lygodactylus picturatus的iota-晶体蛋白与一种改良形式的视觉色素视黄醇结合，以保护其视网膜在明亮阳光下免受紫外线损伤。 核磁共振和光谱、光散射和定向诱变实验将与分子动力学和蒙特卡罗模拟相结合，以阐明赋予这些蛋白质高稳定性和极端条件抵抗力的重要结构因素和分子间相互作用，从而指导仿生材料的未来设计。了解不同晶状体蛋白的共同结构因素将有助于指导未来耐高温和抗紫外线仿生材料的设计。此外，预测和控制蛋白质混合物中液-液相分离的形成和解离的能力将有助于调节蛋白质聚集和生化活性。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Large-Scale Recombinant Production of the SARS-CoV-2 Proteome for High-Throughput and Structural Biology Applications.

• DOI: 10.3389/fmolb.2021.653148
• 发表时间: 2021
• 期刊: Frontiers in molecular biosciences
• 影响因子: 5
• 作者: Altincekic N;Korn SM;Qureshi NS;Dujardin M;Ninot-Pedrosa M;Abele R;Abi Saad MJ;Alfano C;Almeida FCL;Alshamleh I;de Amorim GC;Anderson TK;Anobom CD;Anorma C;Bains JK;Bax A;Blackledge M;Blechar J;Böckmann A;Brigandat L;Bula A;Bütikofer M;Camacho-Zarco AR;Carlomagno T;Caruso IP;Ceylan B;Chaikuad A;Chu F;Cole L;Crosby MG;de Jesus V;Dhamotharan K;Felli IC;Ferner J;Fleischmann Y;Fogeron ML;Fourkiotis NK;Fuks C;Fürtig B;Gallo A;Gande SL;Gerez JA;Ghosh D;Gomes-Neto F;Gorbatyuk O;Guseva S;Hacker C;Häfner S;Hao B;Hargittay B;Henzler-Wildman K;Hoch JC;Hohmann KF;Hutchison MT;Jaudzems K;Jović K;Kaderli J;Kalniņš G;Kaņepe I;Kirchdoerfer RN;Kirkpatrick J;Knapp S;Krishnathas R;Kutz F;Zur Lage S;Lambertz R;Lang A;Laurents D;Lecoq L;Linhard V;Löhr F;Malki A;Bessa LM;Martin RW;Matzel T;Maurin D;McNutt SW;Mebus-Antunes NC;Meier BH;Meiser N;Mompeán M;Monaca E;Montserret R;Mariño Perez L;Moser C;Muhle-Goll C;Neves-Martins TC;Ni X;Norton-Baker B;Pierattelli R;Pontoriero L;Pustovalova Y;Ohlenschläger O;Orts J;Da Poian AT;Pyper DJ;Richter C;Riek R;Rienstra CM;Robertson A;Pinheiro AS;Sabbatella R;Salvi N;Saxena K;Schulte L;Schiavina M;Schwalbe H;Silber M;Almeida MDS;Sprague-Piercy MA;Spyroulias GA;Sreeramulu S;Tants JN;Tārs K;Torres F;Töws S;Treviño MÁ;Trucks S;Tsika AC;Varga K;Wang Y;Weber ME;Weigand JE;Wiedemann C;Wirmer-Bartoschek J;Wirtz Martin MA;Zehnder J;Hengesbach M;Schlundt A
• 通讯作者: Schlundt A

Active Learning Module for Protein Structure Analysis Using Novel Enzymes

使用新型酶进行蛋白质结构分析的主动学习模块

• DOI: 10.35459/tbp.2021.000209
• 发表时间: 2022
• 期刊: The Biophysicist
• 作者: Kelz, Jessica I.;Takahashi, Gemma R.;Safizadeh, Fatemeh;Farahmand, Vesta;Crosby, Marquise G.;Uribe, Jose L.;Kim, Suhn H.;Sprague-Piercy, Marc A.;Diessner, Elizabeth M.;Norton-Baker, Brenna
• 通讯作者: Norton-Baker, Brenna