Revealing Functions for Newly Discovered Proteins by FAST-NMR

描述（由申请人提供）：本研究计划详细说明了我们建立“概念证明”的计划，该计划将支持随后的R01申请，以完善和扩展我们的方法，以加快通过基因组测序鉴定的假设蛋白质的功能和治疗分析。实现这一具有挑战性的目标有望提供有关细胞生物学、发育、进化和生理学的前所未有的丰富信息，这些信息将对人类健康产生重大影响。本申请的目的是进一步验证和提高我们的FAST-NMR（通过核磁共振功能注释筛选技术）测定的效率，为NIH-NIGMS蛋白质结构倡议（PSI）鉴定的假设蛋白质分配功能。FAST-NMR独特地结合了结构生物学，核磁共振配体亲和筛选和生物信息学来发现未知功能蛋白质的生化功能或功能假设。该研究的目标是筛选具有东北结构基因组学联盟（NESG）确定的NMR结构的功能标注和假设蛋白质，以验证FAST-NMR功能分配的准确性。此外，一个高性能亲和色谱-质谱（HPAC-MS）组件将被纳入FAST-NMR筛选，以提高分析的效率和通量。NESG蛋白与生长调节、癌症、抗生素耐药性和其他重要的生物医学靶标有关。获得这些尚未发现的蛋白质的功能分配将为开发治疗各种人类疾病的新疗法提供关键信息。我们的FAST-NMR分析的中心假设是，具有相似功能的蛋白质将具有相似的活性位点结构特征，尽管在序列和结构上存在全局差异。通过识别已知和未知功能蛋白质之间相似的活性位点结构和序列组成，获得功能注释。FAST-NMR是普遍接受的范例的进一步改进，即全局序列或结构同源性推断功能上的相似性。我们提出的研究具有重要意义，因为由此产生的FAST-NMR方法将通过提供一种方法来注释沉积在PDB中功能目前未知的“孤儿”蛋白的数量不断增加，从而解决PSI的重要需求。通过FAST-NMR分析筛选NESG蛋白来鉴定新的治疗靶点对于加快药物发现过程和改善人类健康是非常宝贵的。公共卫生相关性：拟议的研究与人类健康相关，因为加速从基因组测序中鉴定的大量新蛋白质的功能注释将导致发现新的治疗靶点并加快药物发现过程。开发有效的治疗方法对于解决与传染病、癌症和其他疾病有关的全球公共卫生问题至关重要。

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.