Latent Luciferase

潜伏荧光素酶

• 批准号: 9193082
• 负责人: STEPHEN C. MILLER
• 金额: $ 20.94万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-12-15 至 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9193082
• 关键词: Acyl Coenzyme A; Animal Model; Appearance; Applications Grants; Arachidonic Acids; Bacteria; Binding; Biological Assay; Bioluminescence; Carboxylic Acids; Cells; Codon Nucleotides; Coenzyme A Ligases; Detection; Directed Molecular Evolution; Drosophila genus; Enzymes; Evolution; Exploratory/Developmental Grant; Fireflies; Firefly Luciferases; Fluorescence; Gene Expression Profiling; Geometry; Image; In Vitro; Incubated; Insecta; Ligands; Ligase; Light; Luciferases; Mammalian Cell; Mammals; Methods; Modification; Mus; Mutation; Nematoda; Order Coleoptera; Organism; Oxides; Plants; Process; Proteins; Reporter; Reporting; Role; Sequence Homology; Techniques; Testing; Therapeutic; Visceral; Work; adenylate; analog; bioluminescence imaging; design; drug discovery; genetic manipulation; in vivo; light emission; luciferin; non-invasive optical imaging; oxidation; pre-clinical; protein function; protein protein interaction; public health relevance; response; screening; sensor; small molecule

 DESCRIPTION (provided by applicant): Bioluminescence is a powerful, inexpensive, and non-invasive method to monitor gene expression, enzymatic activity, protein-protein interactions and response to therapeutics both in vitro and in vivo. However, bioluminescence requires the use of a luciferase derived from a small number of luminogenic organisms (e.g., fireflies), and is limited to applications and organisms where genetic manipulation is both possible and practical. In this exploratory/developmental grant, we will identify proteins in non-luminous species that are latent luciferases, capable of bioluminescent light emission when treated with an appropriate substrate. In support of this substrate-directed approach, we have identified a fruit fly protein that is revealed to be a luciferase when treated with a synthetic luciferin analog. This unprecedented result shows that the function of a protein can be fundamentally changed simply by modification of the substrate, rather than by mutation. Beyond the paradigm-shifting implications for the evolution and design of new enzymatic activities, the prospect that non-luminogenic organisms encode latent luciferases has transformative potential for the non-invasive optical imaging of endogenous processes, potentially enabling bioluminescence imaging in native cells and organisms without the need for the introduction of an exogenous luciferase.



项目成果

Luciferase Activity of Insect Fatty Acyl-CoA Synthetases with Synthetic Luciferins.

• DOI: 10.1021/acschembio.7b00813
• 发表时间: 2017-12-15
• 期刊: ACS chemical biology
• 影响因子: 4
• 作者: Mofford DM;Liebmann KL;Sankaran GS;Reddy GSKK;Reddy GR;Miller SC
• 通讯作者: Miller SC