Metastable Crystallins: Structure and Stabilization

亚稳态晶体蛋白：结构和稳定性

• 批准号: 9265858
• 负责人: KRISHNA K SHARMA
• 金额: $ 39.17万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9265858
• 关键词: Adult; Aging; Alzheimer' s Disease; Blindness; Body part; Cataract; Cataract Extraction; Cell Culture Techniques; Cells; Client; Complex; Crosslinker; Crystallins; Data; Degenerative Disorder; Deposition; Development; Diabetes Mellitus; Disease; Eye; Glaucoma; Goals; Human; Huntington Disease; Hydrophobicity; Implant; Intervention; Intraocular lens implant device; Investigation; Knowledge; Lead; Learning; Malignant Neoplasms; Methods; Modification; Molecular Chaperones; Mutation; Names; Nuclear; Organ; Parkinson Disease; Pathogenesis; Pathologic; Peptides; Play; Protein Conformation; Proteins; Reagent; Recovery; Research; Role; Savings; Site; Site-Directed Mutagenesis; Structure; Systems Analysis; Testing; Time; Tissues; Treatment Efficacy; age related; alpha-Crystallins; cost; crosslink; experimental study; functional restoration; innovation; insight; lens; lens protein; lens transparency; light scattering; macromolecule; mutant; non-Native; novel; prevent; protein aggregate; protein aggregation; protein misfolding; protein protein interaction; public health relevance; stem; synthetic peptide; tool

DESCRIPTION (provided by applicant): Many diverse diseases, including cataract, some types of glaucoma, Alzheimer disease, diabetes and cancer to name only a few, are now known to share the common feature of aggregated, misfolded or modified protein deposits. The pathological hallmark in this group of diseases is protein aggregation and deposition in specific cells, tissues or organs. The pathological similarities indicate that common principles that govern protein interactions underlie protein misfolding degenerative diseases. Cataract is a classic example of a protein misfolding and aggregation disease. Cataract, a major cause of blindness in the world, develops as a result of age-related modifications and aggregation of the lens proteins. α-crystallin accounts for nearly 40% of the adult lens proteins but its structure-function is yet to be fully understood. The chaperone-like activity of α-crystallin is believed to play a central role in maintaining lens transparency. During aging, lens crystallins undergo truncation and these modifications correlate with lens crystallin aggregations responsible for light scattering. It is well established that the addition of α-crystallin to aggregating proteins stops a further increase in aggregation and light scattering. We have demonstrated that specific sequences in α-crystallin subunits suppress aggregation of denaturing proteins. However, the full potential of α-crystallin-derived peptides (mini-chaperones) is yet to be realized. To enhance our understanding of the role of non-native interactions in cataract formation, we propose the following specific aims: Aim 1. Identify the sequences involved in abnormal interactions of αA-crystallin subunits that lead to protein aggregation in human cataract-causing A-crystallin mutants (αAR49C, αAF71V, αAG98R and αAR116H) using novel deuterated cross linkers, biotynylated reagents and peptide arrays. Aim 2. Determine the mechanism and efficacy of αA-crystallin-derived mini-chaperone in preventing the aggregation of metastable αA- crystallins (mutants and truncated crystallins) and restoring chaperone activity. The specific aims will be accomplished using novel cross-linkers and mass spectrometric methods. We will also use site-directed mutagenesis and cell culture expression and analysis systems to confirm the cross linking data. These innovative studies will give us new insights into potential interventions for protein misfolding diseases, not only of the eye but also of other parts of the body.

描述（由适用提供）：许多潜水疾病，包括CATAACT，某些类型的青光眼，阿尔茨海默氏病，糖尿病和癌症仅举几例，现在已知可以共享聚合，错误折叠或修饰的蛋白质沉积物的共同特征。这组疾病中的病理标志是特定细胞，组织或器官中的蛋白质聚集和沉积。病理相似性表明，控制蛋白质相互作用的常见原理是蛋白质错误折叠疾病的基础。白内障是蛋白质错误折叠和聚集疾病的经典例子。白内障是世界失明的主要原因，是由于年龄相关的修饰和晶状体蛋白的聚集而发展的。 α-晶状体蛋白几乎占成年晶状体蛋白的40％，但其结构功能尚未完全了解。类似伴侣的活性 据信α-晶状体在维持晶状体透明度方面起着核心作用。在衰老过程中，晶状体晶体会经历截断，这些修饰与负责光散射的透镜晶体聚集在一起。众所周知，在聚集蛋白质中添加α-晶蛋白会阻止聚集和光散射的进一步增加。我们已经证明了α-晶状蛋白亚基中的特定序列抑制了变性蛋白的聚集。然而，α-晶链蛋白衍生的肽（迷你链蛋白）的全部潜力尚未实现。增强 Our understanding of the role of non-native interactions in cataract formation, we propose the following specific aims: Aim 1. Identify the sequences involved in abnormal interactions of αA-crystallin subunits that lead to protein aggregation in human cataract-causing A-crystallin mutants (αAR49C, αAF71V, αAG98R and αAR116H) using novel deuterated cross linkers,生物囊化试剂和肽阵列。 AIM 2。确定αA-晶状体衍生的迷你链酮的机制和有效研究，以防止亚稳态的αA-晶状体（突变体和截短的结晶蛋白）和恢复链酮活性的聚集。具体目标将使用新型的交联和质谱法实现。我们还将使用位置定向的诱变和细胞培养表达和分析系统来确认交叉链接数据。这些创新的研究将为我们提供有关蛋白质错误折叠疾病的潜在干预措施的新见解，不仅是眼睛，而且对身体的其他部位。