Design of switchable proteins and enzymes.

可切换蛋白质和酶的设计。

• 批准号: 9135508
• 负责人: STEWART N LOH
• 金额: $ 32.04万
• 依托单位: UPSTATE MEDICAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9135508
• 关键词: Ankyrin Repeat; Antibiotic Resistance; Bacteria; Binding; Binding Proteins; Biochemical; Biological; Biosensor; Cancerous; Cell Proliferation; Cell physiology; Cells; Chimeric Proteins; Complex; Coupling; Detection; Disease; Engineering; Enzymes; FK506; Family; Fibronectins; Fluorescence Resonance Energy Transfer; Generations; Genetic; Goals; Health; Human; In Vitro; Investigation; Ions; Lactamase; Life; Ligand Binding; Link; Mainstreaming; Malignant Neoplasms; Medical; Metallothionein; Metals; Molecular; Nature; Output; Pathway interactions; Pharmaceutical Preparations; Positioning Attribute; Property; Protein Fragment; Proteins; Proteolysis; Receptor Protein-Tyrosine Kinases; Role; Signal Pathway; Signal Transduction; Signaling Protein; Site; Stretching; Tacrolimus Binding Proteins; Technology; Testing; Therapeutic; Time; War; Yeasts; analog; base; design; experience; in vivo; insight; molecular recognition; monomer; novel; protein function; research study; scaffold; screening; sensor; simulation; small molecule; tool

 DESCRIPTION (provided by applicant): This project employs a combined experimental and computational approach to develop mechanisms by which ordinary proteins can be converted into molecular switches. Induced domain swapping (INDOS) is a technology that allows the function of a protein or enzyme to be switched on and off by a small molecule drug, metal ion, or change in pH. INDOS will be used to manipulate cellular pathways, in living cells and in real time, to examine their roles in health and disease. Protein fragment exchange (FREX) and alternate frame folding (AFF) are mechanisms by which an arbitrary binding protein can be turned into a fluorescent biosensor. We will generate a family of sensors for detecting a variety of biological targets in vivo and in vitro. The impact of this study lies in that the switching mechanisms being developed: (i) are general and can be applied to many proteins; (ii) are versatile, allowing the resulting switches to perform a variety of new and useful functions; and (iii) will provide insight into their underlying molecular phenomena (domain swapping, fragment complementation, and circular permutation), all of which occur commonly in nature yet are poorly understood.

 描述（由适用提供）：该项目采用一种组合的实验和计算方法来开发机制，可以将普通蛋白转换为分子开关。诱导结构域交换（Indos）是一项技术，允许通过小分子药物，金属离子或pH变化打开和关闭蛋白质或酶的功能。 Indos将用于在活细胞和实时操纵细胞途径，以检查其在健康和疾病中的作用。蛋白质碎片交换（FREX）和替代框架折叠（AFF）是可以将任意结合蛋白转变为荧光生物传感器的机制。我们将生成一个传感器家族，用于在体内和体外检测各种生物学靶标。这项研究的影响在于开发了开关机制：（i）是一般的，可以应用于许多蛋白质； （ii）是多功能的，允许生成的开关执行各种新的和有用的功能； （iii）将洞悉其潜在的分子现象（域交换，碎片完成和循环排列），所有这些都通常在自然界中出现，但尚未理解。