Functional Ocular Chemoproteomics for Retinal Biology Insight and in vivo Enzyme Activity

用于视网膜生物学洞察和体内酶活性的功能性眼部化学蛋白质组学

基本信息

批准号：
10667228
负责人：
John Douglas Hulleman
金额：
$ 1.24万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10667228
关键词：
Active Sites Affect Affinity Age related macular degeneration Aging Atlases Biological Markers Biology Caspase Cell Separation Cells Chemicals Choroid Communicable Diseases Coupled Data Data Set Development Dimensions Disease Engineering Enzymes Eye Eye diseases Family Genomics Goals Health High temperature of physical object Imaging Techniques Inherited Intervention Knowledge Label Locales Mass Spectrum Analysis Matrix Metalloproteinases Measures Messenger RNA Metabolic Diseases Modification Mus Mutation Neural Retina Normal tissue morphology Outcome Physiologic Monitoring Physiological Proteins Proteome Proteomics Regulation Research Personnel Research Project Grants Retina Risk Factors Scientist Serine Hydrolase Serine Protease Specificity Testing Time Tissues Transcript Validation Vision research activity-based protein profiling age related chemoproteomics cofactor design disorder risk enzyme activity extracellular genome-wide immunocytochemistry in vivo inhibitor insight neural novel protein folding protein function real-time images spatiotemporal tool transcriptome transcriptome sequencing transcriptomics virtual

项目摘要

PROJECT SUMMARY Recent advances in ‘omics-based approaches allow virtually any scientist to perform powerful genome-wide, transcriptome-wide and proteome-wide analyses of normal tissue, diseasedtissue, or even single cells. The information provided by these analyses has afforded important insight into cellular changes that occur as a consequence of development, disease and risk factors associated with disease, and the identification of potential targets for disease intervention. Yet, all too commonly, the vast majority of ‘omics studies enable only inferences regarding the ultimate effect of the observation on protein function. For example, many researchers routinely infer that an increase in abundance either at the transcript or protein level correlates with a corresponding increase in that particular protein’s function; but this is not necessarily the case. In reality, the function of a protein is not simply determined by its mRNA or protein abundance, but rather by the culmination of the cell’s ability to fold the protein appropriately, traffic it to the proper cellular/extracellular locale, and the presence of inhibitors, activators, and/or cofactors. To overcome these inherent limitations of conventional ‘omics approaches, a unique chemical- biology approach called activity-based protein profiling (ABPP) has been used to provide functional protein activity data. Towards this end, in this exploratory/developmental project, we will assess i) the utility of ABPP to determine whether age-related retinal alterations ultimately affect protein function in a physiologic context and ii) whether we can leverage newly designed, enzyme-specific ABPP probes to yield real-time enzyme activity in vivo. Successful completion of this project will positively impact our knowledge of ocular enzymes and will provide additional tools for interrogating the intricate biology underlying incurable age-related and inherited eye diseases.

项目概要 “基于组学的方法”的最新进展几乎使任何科学家都能够发挥强大的作用对正常组织、患病组织进行全基因组、全转录组和全蛋白质组分析，这些分析提供的信息提供了重要的见解。因发育、疾病和风险因素而发生的细胞变化与疾病相关，并确定疾病干预的潜在目标。很常见的是，绝大多数“组学研究”只能对最终结果进行推断例如，许多研究人员通常推断观察结果对蛋白质功能的影响。转录本或蛋白质水平丰度的增加与相应的特定蛋白质的功能增强；但实际上情况并非如此。蛋白质的功能不仅仅由其 mRNA 或蛋白质丰度决定，而是由细胞适当折叠蛋白质、将其运输到适当位置的能力的顶峰细胞/细胞外区域，以及抑制剂、激活剂和/或辅因子的存在。克服传统“组学方法”的这些固有局限性，一种独特的化学方法称为基于活性的蛋白质分析 (ABPP) 的生物学方法已被用来提供为此，在这个探索/开发项目中，我们将评估 i) ABPP 的效用，以确定最终是否会出现与年龄相关的视网膜改变影响生理背景下的蛋白质功能，以及 ii) 我们是否可以利用新设计的，酶特异性 ABPP 探针可产生体内实时酶活性。该项目将对我们对眼酶的了解产生积极影响，并将提供额外的帮助用于探究不可治愈的年龄相关遗传性眼病背后的复杂生物学的工具疾病。