Large-scale mapping of the protein function landscape

蛋白质功能图谱的大规模绘图

• 批准号: 8525538
• 负责人: Philip Anthony Romero
• 金额: $ 4.92万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8525538
• 关键词: Abate; Affect; Amino Acid Sequence; Biological Assay; DNA; DNA Sequence; Data; Disease; Engineering; Enzymes; Evolution; Gene Mutation; Genetic Epistasis; Genomics; Goals; Hereditary Disease; Hour; Human; Human Genetics; Hyperargininemia; In Vitro; Individual; Knowledge; Laboratories; Lead; Malignant Neoplasms; Maps; Medicine; Methods; Microfluidic Microchips; Microfluidics; Modeling; Molecular; Mutation; Nature; Pathway interactions; Peptide Sequence Determination; Property; Protein Engineering; Proteins; Research; Role; Schools; Statistical Models; Technology; Testing; Therapeutic; Variant; arginase; base; chemotherapeutic agent; design; high throughput screening; insight; interest; predictive modeling; protein function; public health relevance; sequence learning; urea cycle

DESCRIPTION (provided by applicant): Knowledge of how mutations affect protein function is important for understanding how natural proteins evolved, engineering new proteins with useful properties, and predicting disease-associated mutations. Epistasis is the phenomenon where the effect of one mutation depends on the presence of another mutation. These epistatic interactions stymie our ability to predict the phenotypic consequences of mutations, and can restrict the mutational pathways that are available to evolution. Laboratory evolution studies have highlighted the multifaceted nature of protein function and how competing biophysical properties, such as enzymatic activity and protein stability, can generate epistatic interactions between residues. I hypothesize that most epistasis arises as a result of these pleiotropic mechanisms, rather than direct physical interactions between residues. The human urea cycle enzyme Arginase I (hArgI) will be used to study how sequence changes affect expression, stability, and enzymatic activity. The experimental approach will leverage recent advances in parallel DNA sequencing and ultra-high- throughput screening to map the protein function landscape on an unprecedented scale. These data will be used to explore how combinations of biophysical properties generate mutational epistasis and define the space of functional protein sequences.

描述（由申请人提供）：了解突变如何影响蛋白质功能对于理解天然蛋白质如何进化、设计具有有用特性的新蛋白质以及预测与疾病相关的突变非常重要。上位性是一种突变的影响取决于另一种突变的存在的现象。这些上位相互作用阻碍了我们预测突变表型后果的能力，并且可以限制可用于进化的突变途径。实验室进化研究强调了蛋白质功能的多方面性质，以及酶活性和蛋白质稳定性等竞争性生物物理特性如何在残基之间产生上位相互作用。我假设大多数上位性是这些多效性机制的结果，而不是残基之间的直接物理相互作用。人类尿素循环酶精氨酸酶 I (hArgI) 将用于研究序列变化如何影响表达、稳定性和酶活性。该实验方法将利用并行 DNA 测序和超高通量筛选的最新进展，以前所未有的规模绘制蛋白质功能图谱。这些数据将用于探索生物物理特性的组合如何产生突变上位性并定义功能蛋白序列的空间。