New Tools to Measure and Correct Endoplasmic Reticulum Stress in Single Living

测量和纠正单身生活内质网应力的新工具

基本信息

批准号：
7429340
负责人：
Feroz R Papa
金额：
$ 231.63万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
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：
2007-09-30 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7429340
关键词：
26S proteasome 3&apos Untranslated Regions Address Adenosine Adenoviruses Affect Alleles Animal Model Animals Antibodies Apoptosis Apoptotic Appointment Autoimmunity Award B-Lymphocytes Basic Science Behavior Binding Biochemical Biochemistry Biogenesis Biological Assay Biological Factors Biological Sciences Biology Biomedical Research Bite Buffers Bypass Calcium California Cell Death Cell Differentiation process Cell Line Cell Nucleus Cell Separation Cell Transplants Cell secretion Cells Cellular biology Cessation of life Chicago Class Cleaved cell Client Clinic Clinical Clinical Endocrinology Clinical Medicine Co-Immunoprecipitations Collaborations Collection Color Complex Condition Consumption County Crowding Cultured Cells Cysteine Cytoprotection Cytosol Data Development Diabetes Mellitus Dimensions Disease Disruption Disulfide Linkage Disulfides Doctor of Philosophy Dose Drug Delivery Systems Drug usage Educational process of instructing Electron Transport Electrons Embryo Endoplasmic Reticulum Endoribonucleases Energy Transfer Engineering Ensure Environment Enzyme-Linked Immunosorbent Assay Enzymes Essential Genes Eukaryota Eukaryotic Cell Evaluation Event Experimental Designs Face Faculty Failure Fibroblasts Figs - dietary Flow Cytometry Fluorescence Friends Fright Functional disorder Funding Future G Cells Gel Gene Deletion General Hospitals Generations Genes Genetic Genetic Screening Genetic Transcription Genome Glucose Glutathione Disulfide Goals Grant Gray unit of radiation dose Green Fluorescent Proteins Hand Harvest Health Homeostasis Hormones Housing Human Illinois Immunoglobulins In Situ Nick-End Labeling In Vitro Indium Individual Infection Institutes Institution Insulin Insulin Resistance Integral Membrane Protein Islet Cell Islets of Langerhans Islets of Langerhans Transplantation Killer Cells Knowledge Label Lasers Lead Learning Left Life Ligand Binding Ligands Link Localized MAPK8 gene Maintenance Malignant Neoplasms Mammals Maps Mass Spectrum Analysis Measurable Measures Mediating Medical Membrane Membrane Proteins Mentors Messenger RNA Methodology Metric Microarray Analysis Microscopy Missense Mutation Mission Modeling Modification Molecular Molecular Chaperones Monitor Morus Multiple Myeloma Mus Mutagenesis Mutation Nature Neoplasm Metastasis Nerve Degeneration Noise Non-Insulin-Dependent Diabetes Mellitus Nucleotides Numbers Nutrient Obesity Occupations One-Step dentin bonding system Optics Oranges Organelles Organism Orthologous Gene Outcome Output Oxidation-Reduction Oxidative Stress Oxidoreductase Oxygen Pancreas Pancreatic ribonuclease Paper Pathogenesis Pathology Pathway interactions Patients Peripheral Pharmaceutical Preparations Phosphotransferases Physicians Physiologic pulse Physiological Physiology Plasma Plasma Cells Population Positioning Attribute Postdoctoral Fellow Prize Procedures Process Production Proinsulin Property Protein Disulfide Isomerase Protein Engineering Protein Glycosylation Protein Kinase Protein Overexpression Protein Secretion Proteins Protocols documentation Psychological reinforcement Pulse taking Purpose Quality Control RNA RNA Interference RNA Splicing Range Rate Rattus Reactive Oxygen Species Recombinant Proteins Recombinants Reduced Glutathione Regulation Relative (related person)Reporter Reporting Research Research Personnel Resistance Resources Response Elements Rest Ribonucleases Risk Rodent Role Running S Phase Saccharomyces cerevisiae Saccharomycetales San Francisco Science Score Secretory Cell Shapes Side Signal Pathway Signal Transduction Small Interfering RNA Solid Solutions Sorting - Cell Movement Source Specificity Staging Staining method Stains Standardization Standards of Weights and Measures Steam Stress Structure Structure-Activity Relationship Students Surface Syndrome System Tail Techniques Technology Testing Tetanus Helper Peptide Text Thinking Time Training Transducers Translations Transplantation Traumatic Stress Disorders Triage Tunicamycin Ubiquitin United States National Institutes of Health Universities Upper arm Variant Viola Work Writing Yeasts analog annexin A5 base biological adaptation to stress cDNA Arrays cancer cell cell transformation cell type cellular engineering chemical genetics chemical reduction chimeric gene college comparative concept cost cytotoxicity design desire diabetic disulfide bond drug development experience falls fascinate fluorophore gene delivery system gene function glycosylation human disease improved in vitro Assay in vivo inhibitor/antagonist innovation insulinoma interest islet knock-down mathematical model member mutant novel oxidation pandemic disease paralogous gene peer periplasm polypeptide preclinical study preference pressure prevent professor programs protein aggregation protein folding protein misfolding reconstitution release of sequestered calcium ion into cytoplasm repaired research study response secretory protein sensor size small molecule small molecule libraries sound success sugar synthetic protein time use tool transcription factor two-dimensional

项目摘要

Aided by chaperones and other activities, proteins of the secretory pathway fold to their correct shapes in the endoplasmic reticulum (ER). But, if ER folding capacity is exceeded, unfolded proteins start to aggregate. This imbalanced condition—called ER stress—is being linked to diverse diseases, such as type 2 diabetes and cancer. The unfolded protein response (UPR) can rebalance a stressed ER, but if the stress is too great (or the response too weak), cells appear to cross a “tipping point”, and undergo apoptosis. This could explain why overworked insulin-producing pancreatic β-cells die in type 2 diabetes. On the other hand, a hyperactive UPR may allow malignant cells to survive hostile environments, promoting cancer. If we could measure both ER stress and the strength of corrective responses directly in healthy and diseased cells, we would be able to test these ideas quantitatively, and design rational therapies for these diseases. Structural engineers study system integrity by applying loads, measuring stresses, perturbing reinforcements, and making corrections. We propose similar approaches to ER stress disorders. Protein secretion generates and consumes oxidizing equivalents, so it should be possible to follow deviations in the ER's redox potential from its resting setpoint as an analog measure of ER stress. For this purpose, we are making redox-responsive fluorescent proteins to gauge the ER's redox potential in single, living cells. We have learned to stably perturb ER protein folding by converting an unfolded protein sensor called Ire1 into a finely adjustable rheostat. In living pancreatic β-cells, these tools will allow us to quantify the ER stress caused by insulin folding load, in both healthy and diabetic states. Finally, using a biochemical assay we have developed, we will discover molecules targeting Ire1, to increase or decrease ER folding ability, as appropriate corrective measures for various ER stress disorders.

在伴侣和其他活动的帮助下，分泌途径的蛋白质折叠在内质网 (ER) 中形成正确的形状但是，如果 ER 折叠。超出容量，未折叠的蛋白质开始聚集。称为 ER 应激的病症与多种疾病有关，例如 2 型疾病未折叠蛋白反应（UPR）可以重新平衡糖尿病和癌症。强调内质网，但如果压力太大（或反应太弱），细胞似乎跨越了“临界点”，并发生细胞凋亡，这可以解释为什么。过度劳累的产生胰岛素的胰腺 β 细胞会在 2 型糖尿病中死亡。另一方面，过度活跃的 UPR 可能使恶性细胞能够在敌对环境中存活下来。如果我们能够同时测量内质网压力和内质网压力，就会促进癌症的发生。直接在健康和患病细胞中的纠正反应的强度，我们会能够定量地测试这些想法，并为这些想法设计合理的疗法疾病。结构工程师通过施加载荷、测量来研究系统完整性我们提出类似的建议。蛋白质分泌产生和消耗的方法。氧化当量，因此应该可以跟踪 ER 中的偏差其静息设定点的氧化还原电位作为 ER 应力的模拟量度。为此，我们正在制造氧化还原响应荧光蛋白来测量单个活细胞中 ER 的氧化还原电位我们已经学会了稳定地扰动 ER。通过将称为 Ire1 的未折叠蛋白质传感器转换为精细的蛋白质折叠在活的胰腺 β 细胞中，这些工具将使我们能够量化健康和糖尿病患者中胰岛素折叠负荷引起的 ER 应激最后，使用我们开发的生化检测，我们将发现。靶向 Ire1 的分子，以增加或减少 ER 折叠能力，如针对各种 ER 应激障碍采取适当的纠正措施。