Revealing Functions for Newly Discovered Proteins by FAST-NMR

描述(由申请人提供)：本研究建议书详细说明了我们建立“概念证明”的计划，该计划将支持后续的R01申请，以完善和扩展我们的方法学，以加快通过基因组测序识别的假想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关于细胞生物学、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。这项应用的目的是进一步验证和提高我们的快速核磁共振(FAST-核磁共振功能注释筛选技术)分析的效率，以将功能分配给NIH-NIGMS蛋白质结构倡议(PSI)确定的假设蛋白质。快速核磁共振将结构生物学、核磁共振配基亲和筛选和生物信息学相结合，发现未知功能蛋白质的生化功能或功能假说。这项研究的目标是筛选具有东北结构基因组联合会(NESG)确定的核磁共振结构的功能注释和假设的蛋白质，以验证快速核磁共振功能分配的准确性。此外，高效亲和层析-质谱仪(HPAC-MS)组件将被整合到快速核磁共振筛选中，以提高分析的效率和吞吐量。Nesg蛋白与生长调节、癌症、抗生素耐药性和其他生物医学上的重要靶点有关。到目前为止，获得这些尚未发现的蛋白质的功能分配应该为开发治疗各种人类疾病的新疗法提供关键信息。我们快速核磁共振分析的中心假设是，尽管全球在序列和结构上存在差异，但功能相似的蛋白质将具有相似的活性部位结构特征。通过识别已知和未知功能的蛋白质之间相似的活性部位结构和序列组成，获得功能注释。快速核磁共振是普遍接受的范例的进一步改进，即全球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，因为由此产生的快速核磁共振方法将通过提供一种方法来注释沉积在PDB中的功能目前未知的越来越多的“孤儿”蛋白质，从而满足PSI的一个重要需求。在我们的快速核磁共振分析中，通过筛选Neng蛋白来识别新的治疗靶点，对于加快药物发现过程和改善人类健康具有非常重要的价值。公共卫生相关性：拟议的研究与人类健康相关，因为加快从基因组测序中识别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、癌症和其他疾病相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.