Revealing Functions for Newly Discovered Proteins by FAST-NMR

通过 FAST-NMR 揭示新发现蛋白质的功能

基本信息

批准号：
7828040
负责人：
ROBERT POWERS
金额：
$ 17.84万
依托单位：
UNIVERSITY OF NEBRASKA LINCOLN
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-05-07 至 2012-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7828040
关键词：
Active Sites Address Affinity Affinity Chromatography Antibiotic Resistance Biochemical Bioinformatics Biological Biological Assay Biological Process Cancer cell line Cellular biology Characteristics Communicable Diseases Communities Data Set Deposition Development Evolution Family Genomics Goals Growth Health Human Infectious Agent Lead Left Ligand Binding Ligands Malignant Neoplasms Mass Spectrum Analysis Methodology Orphan PAWR protein Performance Pharmaceutical Preparations Physiology Process Protein Structure Initiative Proteins Proteome Public Health Regulation Research Research Proposals Role Screening procedure Structure Technology Therapeutic United States National Institutes of Health design drug discovery functional genomics human disease improved innovation interest microbial new therapeutic target novel novel strategies pathogen protein complex public health relevance structural biology structural genomics therapeutic target three dimensional structure

项目摘要

DESCRIPTION (provided by applicant): This research proposal details our plan to establish the "proof of concept" that will support a subsequent R01 application to perfect and extend our methodology to expedite the functional and therapeutic analysis of hypothetical proteins identified by genomic sequencing. Accomplishing this challenging goal promises to provide an unprecedented wealth of information about cell biology, development, evolution and physiology that will have a significant impact on human health. The aim of this application is to further validate and increase the efficiency of our FAST-NMR (functional annotation screening technology by NMR) assay to assign a function to hypothetical proteins identified by the NIH-NIGMS Protein Structure Initiative (PSI). FAST-NMR uniquely combines structural biology, NMR ligand affinity screens and bioinformatics to discover biochemical functions or functional hypotheses of proteins of unknown function. The goal of the proposed research is to screen functionally annotated and hypothetical proteins with NMR structures determined by the Northeast Structural Genomics Consortium (NESG) to verify the accuracy of FAST-NMR functional assignments. Additionally, a high-performance affinity chromatography-mass spectrometry (HPAC-MS) component will be incorporated into the FAST-NMR screen to improve the efficiency and throughput of the assay. NESG proteins are associated with growth regulation and cancer, antibiotic resistance and other biomedically-important targets. Obtaining a functional assignment for these as of yet undiscovered proteins should provide key information for developing new therapies to treat various human diseases. The central hypothesis of our FAST-NMR assay is that proteins with similar function will have similar active-site structural characteristics, despite global differences in sequence and structure. A functional annotation is obtained by identifying similar active-site structure and sequence composition between proteins of known and unknown function. FAST-NMR is a further refinement of the generally accepted paradigm that global sequence or structure homology infers a similarity in function. Our proposed research is significant because the resultant FAST-NMR methodology will address an important need of PSI by providing an approach to annotate the expanding number of "orphaned" proteins deposited in the PDB whose function are currently unknown. The identification of novel therapeutic targets by screening NESG proteins in our FAST-NMR assay is invaluable for expediting the drug discovery process and improving human health. PUBLIC HEALTH RELEVANCE: The proposed research is relevant to human health because accelerating the functional annotation of the vast number of novel proteins identified from genomic sequencing will lead to the discovery of new therapeutic targets and expedite the drug discovery process. The development of effective therapeutics is essential to addressing global public health problems associated with infectious disease, cancer and other illnesses.

描述（由申请人提供）：本研究建议详细介绍了我们建立“概念证明”的计划，该计划将支持随后的R01应用，以完善和扩展我们的方法，以加快基因组测序鉴定的假设蛋白的功能和治疗分析。实现这个挑战性的目标有望提供有关细胞生物学，开发，进化和生理学的空前信息，这将对人类健康产生重大影响。该应用的目的是进一步验证和提高我们的快速NMR（通过NMR功能注释筛选技术）测定的效率，以将功能分配给NIH-NIGMS蛋白质结构Initiative（PSI）确定的假设蛋白。 Fast-NMR独特地结合了结构生物学，NMR配体亲和力筛选和生物信息学，以发现未知功能的蛋白质的生化功能或功能假设。拟议的研究的目的是用由东北结构基因组学联盟（NESG）确定的NMR结构进行功能注释和假设的蛋白质，以验证快速NMR功能分配的准确性。此外，将将高性能亲和力色谱 - 质谱法（HPAC-MS）组件纳入快速-NMR屏幕中，以提高测定法的效率和吞吐量。 NESG蛋白与生长调节和癌症，抗生素耐药性和其他生物医学重要靶标有关。从未发现的蛋白质获得功能分配，应为开发新的治疗各种人类疾病的新疗法提供关键信息。我们快速NMR测定法的中心假设是，尽管序列和结构的全球差异，但具有相似功能的蛋白质将具有相似的活动位置结构特征。通过识别已知功能和未知功能的蛋白质之间的相似的活性位点结构和序列组成来获得功能注释。 Fast-NMR是对全局序列或结构同源性功能相似性的普遍接受范式的进一步完善。我们提出的研究很重要，因为最终的快速NMR方法将通过提供一种方法来注释存放在功能目前未知的PDB中的“孤儿”蛋白质扩大的“孤儿”蛋白质来满足PSI的重要需求。通过在我们的快速NMR分析中筛选NESG蛋白来鉴定新的治疗靶标，对于加快药物发现过程并改善人类健康是无价的。公共卫生相关性：拟议的研究与人类健康有关，因为从基因组测序中鉴定出的大量新型蛋白质的功能注释将导致发现新的治疗靶标，并加快药物发现过程。有效治疗的发展对于解决与传染病，癌症和其他疾病有关的全球公共卫生问题至关重要。