Design of switchable proteins and enzymes.

可切换蛋白质和酶的设计。

基本信息

批准号：
9301601
负责人：
STEWART N LOH
金额：
$ 31.85万
依托单位：
UPSTATE MEDICAL UNIVERSITY
依托单位国家：
美国
项目类别：
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/9301601
关键词：
Ankyrin Repeat Antibiotic Resistance Bacteria Binding Binding Proteins Biochemical Biological Biophysics Biosensor Biotechnology Cancerous Cell Proliferation Cell physiology Cells Chimeric Proteins Complement Complex Coupling Detection Dimerization Disease Engineering Enzymes FK506 Family Fibronectins Fluorescence Resonance Energy Transfer Generations Genetic Goals HRAS gene Health Human In Vitro Investigation Ions Lactamase Ligand Binding Link Mainstreaming Malignant Neoplasms Medical Metallothionein Metals Molecular Molecular Conformation Nature Output Pathway interactions Pharmaceutical Preparations Phenotype 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 Thermodynamics Time War Yeasts analog base design dimer experience experimental study in vivo insight molecular recognition monomer novel protein function public health relevance scaffold screening sensor simulation small molecule tool

项目摘要

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（由申请人提供）：该项目采用实验和计算相结合的方法来开发将普通蛋白质转化为分子开关的机制。诱导结构域交换（INDOS）是一种允许蛋白质或酶的功能发生变化的技术。通过小分子药物、金属离子或 pH 值的变化来打开和关闭的 INDOS 将用于实时操纵活细胞中的细胞通路，以检查它们在健康和疾病中的作用。和交替框架折叠（AFF）是一种将任意结合蛋白转变为荧光生物传感器的机制，我们将产生一系列传感器，用于检测体内和体外的各种生物靶标。正在开发的开关机制：（i）是通用的，可以应用于许多蛋白质；（ii）是通用的，允许产生的开关执行各种新的和有用的功能；（iii）将提供对其潜在分子现象的深入了解；（结构域交换、片段互补和循环排列），所有这些在自然界中都很常见，但人们却知之甚少。