Department of Biochemistry, University of Illinois, Urbana, Illinois 61801, USA.
Biochemistry. 2011 Nov 22;50(46):9950-62. doi: 10.1021/bi201312u. Epub 2011 Oct 26.
The Enzyme Function Initiative (EFI) was recently established to address the challenge of assigning reliable functions to enzymes discovered in bacterial genome projects; in this Current Topic, we review the structure and operations of the EFI. The EFI includes the Superfamily/Genome, Protein, Structure, Computation, and Data/Dissemination Cores that provide the infrastructure for reliably predicting the in vitro functions of unknown enzymes. The initial targets for functional assignment are selected from five functionally diverse superfamilies (amidohydrolase, enolase, glutathione transferase, haloalkanoic acid dehalogenase, and isoprenoid synthase), with five superfamily specific Bridging Projects experimentally testing the predicted in vitro enzymatic activities. The EFI also includes the Microbiology Core that evaluates the in vivo context of in vitro enzymatic functions and confirms the functional predictions of the EFI. The deliverables of the EFI to the scientific community include (1) development of a large-scale, multidisciplinary sequence/structure-based strategy for functional assignment of unknown enzymes discovered in genome projects (target selection, protein production, structure determination, computation, experimental enzymology, microbiology, and structure-based annotation), (2) dissemination of the strategy to the community via publications, collaborations, workshops, and symposia, (3) computational and bioinformatic tools for using the strategy, (4) provision of experimental protocols and/or reagents for enzyme production and characterization, and (5) dissemination of data via the EFI's Website, http://enzymefunction.org. The realization of multidisciplinary strategies for functional assignment will begin to define the full metabolic diversity that exists in nature and will impact basic biochemical and evolutionary understanding, as well as a wide range of applications of central importance to industrial, medicinal, and pharmaceutical efforts.
酶功能倡议(EFI)最近成立,旨在解决为细菌基因组项目中发现的酶分配可靠功能的挑战;在本期专题中,我们回顾了 EFI 的结构和运作。EFI 包括超级家族/基因组、蛋白质、结构、计算和数据/传播核心,为可靠预测未知酶的体外功能提供了基础设施。功能分配的初始目标是从五个功能多样的超级家族(酰胺水解酶、烯醇酶、谷胱甘肽转移酶、卤代烷酸脱卤酶和异戊二烯合成酶)中选择,其中五个超级家族特定的桥梁项目通过实验测试了预测的体外酶活性。EFI 还包括微生物学核心,用于评估体外酶功能的体内背景,并确认 EFI 的功能预测。EFI 向科学界提供的成果包括:(1) 开发大规模的、多学科的基于序列/结构的功能分配策略,用于基因组项目中发现的未知酶(目标选择、蛋白质生产、结构确定、计算、实验酶学、微生物学和基于结构的注释),(2) 通过出版物、合作、研讨会和座谈会向社区传播该策略,(3) 用于使用该策略的计算和生物信息学工具,(4) 提供酶生产和表征的实验方案和/或试剂,以及(5) 通过 EFI 的网站 http://enzymefunction.org 传播数据。功能分配的多学科策略的实现将开始定义自然界中存在的全部代谢多样性,并将影响基础生化和进化理解,以及对工业、医学和制药努力至关重要的广泛应用。