COLLABORATIVE CENTER FOR AN ENZYME FUNCTION INITIATIVE

描述（由申请人提供）：酶功能倡议（Enzyme Function Initiative，简称EFP）将开发一种基于序列/结构的稳健策略，以促进发现基因组计划中发现的未知酶的体外酶促和体内代谢/生理功能，这是基因组生物学的一个关键限制。该中心将通过整合生物信息学、结构生物学和计算与酶学、遗传学和代谢组学来实现这一目标。 该中心将建立五个科学核心：1）指导靶点选择，并根据序列关系和基因组背景设计功能分配策略; 2）靶点的表达和纯化; 3）靶点结构的实验测定; 4）目标结构的计算确定（同源性建模），以及配体文库的计算机对接，以通过聚焦文库筛选指导体外功能的实验分配;和5）体外指定功能的体内作用的微生物学和代谢组学表征。 功能预测将测试的五个桥接项目，侧重于功能多样的酰胺水解酶（AH），烯醇化酶（EN），谷胱甘肽转移酶（GST），卤代烷酸脱卤酶（HAD），和类异戊二烯合酶（IS）超家族。选择这些超家族是因为不能仅基于序列或结构相似性通过转移先前的注释来完成功能分配：每个超家族内的反应共享保守的部分反应，但底物/产物的身份不保守。 该中心将向科学界传播知识，计算和实验工具，协议，材料和指导方针，用于确定未知酶的体外和体内功能。在实现这一目标的过程中，基因组计划将使一个更大的研究者联盟成为核心，并使其能够实现基因组计划提供的大量序列数据的生物医学潜力。

DESCRIPTION (provided by applicant): The Enzyme Function Initiative (EFI) will develop a robust sequence/structure-based strategy for facilitating discovery of in vitro enzymatic and in vivo metabolic/physiological functions of unknown enzymes discovered in genome projects, a crucial limitation in genomic biology. The EFI will accomplish this goal by integrating bioinformatics, structural biology, and computation with enzymology, genetics, and metabolomics. The EFI will establish five Scientific Cores for: 1) directing target selection as well as devising strategies for functional assignment based on sequence relationships and genome context; 2) expression and purification of targets; 3) experimental determination of structures of targets; 4) computational determination of structures of targets (homology modeling) and, also, in silico docking of ligand libraries to direct experimental assignment of in vitro functions by focused library screening; and 5) microbiological and metabolomic characterization of the in vivo roles of the in vitro assigned functions. The functional predictions will be tested by five Bridging Projects that focus on the functionally diverse amidohydrolase (AH), enolase (EN), glutathione transferase (GST), haloalkanoic acid dehalogenase (HAD), and isoprenoid synthase (IS) superfamilies. These superfamilies were selected because functional assignment cannot be accomplished by transfer of prior annotations based only on sequence or structural similarity: the reactions within each superfamily share conserved partial reactions but the identities of the substrates/products are not conserved. The EFI will disseminate to the scientific community the intellectual, computational, and experimental tools, protocols, materials, and guidelines for determining in vitro and in vivo functions of unknown enzymes. In achieving this goal, the EFI will nucleate and enable a larger consortium of investigators working toward the goal of realizing the biomedical potential of the vast amount of sequence data provided by genome projects.