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Mechanisms of Activation for Human Small Heat Shock Proteins: An Integrated Approach

人类小热休克蛋白的激活机制：综合方法

基本信息

批准号：
9304219
负责人：
Rachel E Klevit
金额：
$ 61.53万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-05-01 至 2020-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9304219
关键词：
Affect Affinity Age related macular degeneration Binding Biological Assay Blindness Brain Hypoxia-Ischemia Cataract Cell Aging Cells Cellular Stress Charge Chronic Client Complex Cornea Crystalline Lens Detection Diabetic Retinopathy Dimerization Disease Emerging Technologies Equilibrium Exhibits Eye Fluorescence Resonance Energy Transfer Functional disorder Goals HSPB1 gene Health Heat shock proteins Heat-Shock Response Histidine Human Inherited Ions Knowledge Lead Link Maintenance Mammals Mass Spectrum Analysis Metal Ion Binding Metals Modeling Molecular Molecular Chaperones Mutation Names National Eye Institute Negative Staining Neurodegenerative Disorders Oxidative Stress Pathology Physiological Play Property Proteins Refractory Regulation Resolution Retina Role Stress Structural Models Structure Temperature Testing Tissues UV Radiation Exposure Work Zinc base dimer disease phenotype divalent metal dynamic system emergency service responder experience experimental study insight lens monomer protein aggregation protein function protein misfolding protein structure public health relevance response solid state nuclear magnetic resonance structural biology

项目摘要

DESCRIPTION (provided by applicant): Cells have numerous strategies to cope with the consequences of stresses that cause protein misfolding and aggregation, leading to formation of plaques, fibrils, and other aggregated species encountered in aging cells, cataract, and neurodegenerative diseases. The protein chaperones known as small heat shock proteins are the cell's first responders and are therefore key to maintenance of cellular health. Despite their name, human small heat shock proteins (sHSP) are rarely called up to respond to the stress of elevated temperature, as humans have effective and tight regulation of temperature. Nevertheless, much of the current knowledge of sHSPs has been obtained under elevated temperature conditions where they are highly active. In this project, we seek to understand the molecular mechanisms by which human sHSPs respond to physiologically relevant stresses and how they maintain cellular proteins in soluble forms. Of relevance to the National Eye Institute, the sHSPs to be focused on are highly expressed in ocular tissues (lens, retina, cornea) as well as in other tissues. Dysfunction and aberrant expression of HSPB1 and HSPB5 are associated with ocular diseases including cataract, diabetic retinopathy, and age-related macular degeneration that together account for 65% of blindness worldwide. Perhaps linked to their particular functional niche in which they must respond to small changes in cellular conditions such as pH and metal ion concentrations, human sHSPs have evolved properties that are unique in the protein world. They exist as large (> 100 kDa) polydisperse and dynamic assemblies that defy conventional structural biology approaches. We will apply emerging technologies capable of providing molecular and possible atomic-level information about heterogeneous and dynamic systems. Structural information obtained from solution- and solid-state NMR, negative-stain and cryo-EM, and native mass spectrometry will be integrated with dynamic information from FRET-based subunit dynamics and NMR and with functional information from activity assays and client-binding experiments to define the mechanism(s) by which HSPB5 and HSPB1 are activated by conditions brought about by stresses relevant to tissues of the eye such as hypoxia, ischemia, and UV exposure. Both wild-type sHSPs and forms carrying inherited disease mutations will be investigated.