权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Molecular mechanisms of alkane hydroxylase (AlkB) reactivity and selectivity

烷烃羟化酶 (AlkB) 反应性和选择性的分子机制

基本信息

批准号：
10671699
负责人：
RACHEL Narehood AUSTIN
金额：
$ 29.27万
依托单位：
BARNARD COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-15 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10671699
关键词：
Acceleration Active Sites Affect Alcohols Alkane 1-monooxygenase Alkanes Amino Acid Sequence Architecture Atmosphere Attention Deficit Disorder Bacteria Behavior Binding Biochemical Biological Biological Assay Biological Models Bioremediations Biotechnology Carboxylic Acids Chemistry Chimeric Proteins Comparative Study Complex Computer Simulation Development Diabetes Mellitus Electron Transport Elements Engineering Environment Enzymes Escherichia coli Eukaryotic Cell Family Fatty Acid Desaturases Fatty Acids Future Gram-Positive Bacteria Health Homologous Gene Human Hydrogen Bonding Iron Knowledge Learning Length Life Link Lipids Malignant Neoplasms Maps Mediating Membrane Membrane Proteins Metabolism Mixed Function Oxygenases Modeling Molecular Nature Nerve Degeneration Nitrogen Obesity Oils Oxidation-Reduction Oxygen Oxygenases Pathogenicity Pathway interactions Physiological Play Positioning Attribute Process Protein Family Proteins Reaction Red Algae Research Personnel Resolution Role Route Rubredoxins Scientist Structure Substrate Specificity Testing Therapeutic Work Zinc austin biophysical analysis chemical bond clinically relevant computer studies experimental study insight member metalloenzyme mutant novel overexpression oxidized lipid protein folding rapid test spectroscopic survey therapeutic enzyme therapeutic target three dimensional structure

项目摘要

Project Summary Reactions with atmospheric oxygen are required for many life-sustaining processes. The class-III diiron proteins use oxygen to selectively oxidize lipids and to put OH groups into molecules in critical biosynthetic pathways. Class-III diiron enzymes play essential roles in many aspects of lipid synthesis and metabolism and are linked to human health problems including obesity, diabetes, attention-deficit disorder, and neurodegeneration. They are also crucial in the natural bioremediation of oil. There is a dearth of mechanistic information about this family of membrane enzymes, primarily because their membrane-associated nature makes them very difficult to purify and study. Alkane monooxygenase (AlkB) is a member of the class-III integral membrane diiron proteins along with fatty acid desaturases and fatty acid hydroxylases. The amino acid sequence of AlkB indicates that it is not structurally similar to other enzymes with similar functions. Determining its three-dimensional structure is a feat that has eluded scientists for decades. In an important step forward in preliminary work, PI Austin and co-Investigator Feng have solved the first structure of AlkB with a bound substrate and shown that it serves as an excellent model system to understand the catalytic mechanism of class-III diiron proteins. This breakthrough, together with the establishment of a novel assay for rapid functional characterization and the development of a suite of AlkB active AlkB homologs, paves the way to answering key questions about these important metalloenzymes. The PIs will integrate structural, functional, biochemical, computational, and spectroscopic studies to determine the three-dimensional structure of the diiron active site, identify determinants of substrate specificity, learn how AlkB is activated by its partner protein, and probe how the presence of a covalently bound electron-transfer partner, found only in a class of gram positive bacteria, changes the reactivity of this enzyme family. In so doing, they will expand the basic knowledge of strategies to break and make key chemical bonds, which may lead to the development of new synthetic routes to make life-saving and life-extending molecules. Their work will also provide critical insights to efforts to target this family of enzymes for therapeutic purposes.

项目总结