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Defining the conformational control of nitric oxide synthases by a multipronged approach

通过多管齐下的方法定义一氧化氮合酶的构象控制

基本信息

批准号：
10621327
负责人：
Changjian Feng
金额：
$ 31.42万
依托单位：
UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
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：
2020-08-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10621327
关键词：
Alzheimer&apos s Disease Anabolism Architecture Back Behavior Binding Biochemical Calmodulin Complex Computer Analysis Computer Models Dependence Deuterium Development Disease Docking Electron Spin Resonance Spectroscopy Electron Transport Electrons Enzymes Family Flavin Mononucleotide Free Energy Genetic Code Health Heart Heme Hemeproteins Human Hydrogen Kinetics Knowledge Mass Spectrum Analysis Measurement Methods Molecular Molecular Conformation Motion NOS3 gene Neurons Nitric Oxide Nitric Oxide Synthase Output Phosphorylation Phosphoserine Physiologic pulse Post-Translational Regulation Production Property Protein Conformation Protein Dynamics Protein Isoforms Proteins Regulation Research Shapes Site Spectrum Analysis Stimulus Stroke Structure System Techniques Testing Work computational basis effective therapy experimental study flexibility improved in vivo innovation milligram novel therapeutic intervention novel therapeutics programs rate of change response single-molecule FRET statistics synergism two-dimensional

项目摘要

Project Summary The neuronal & endothelial nitric oxide (NO) synthase (nNOS & eNOS) enzymes make NO in response to calmodulin (CaM) binding, and function broadly in human health and disease. Posttranslational regulation through phosphorylation further regulates NOS in vivo in response to stimuli. Hallmarks of these large flavo- hemoproteins include multi-domain architecture with flexible linkers, allowing for dynamic, regulated interdomain electron transfer (IET). NO synthesis requires a large conformational change, in which the FMN domain shuttles between NOS's electron-accepting “input state” and electron-donating “output state” to deliver electrons across the domains. These large-scale motions are shaped by conformational energy landscape, i.e., the dependence of free energy on protein conformation. Moreover, local conformational adjustment likely continues in the docked state. Despite extensive research efforts, the dynamics underlying these conformational changes required for IET across the NOS domains remain unclear. A roadblock to answering this central question is the lack of a unified theoretical/computational approach to interpret the experimental results quantitatively. Solving this vexing research problem calls for a convergence of mesoscopic computational analysis and hands-on experiments that are sensitive to NOS protein dynamics in solution. Combining these latest experimental methods in a multipronged effort is innovative, as it dramatically expands the overall scope of the experimental measurements and provides a better basis for the computations. This approach will allow us to interpret the diverse experimental results and apply them to the calculated NOS conformational behavior paradigm in a consistent manner. Our integrated program draws on the unique combined expertise of the collaborative team. Importantly, we have made the crucial first step of implementing our experimental and computational approaches synergistically. To determine the energy landscape and the resulting NOS conformational properties, we will first calculate the conformational statistics and dynamics and use it in synergy with the suitable experiments to study long-range tethered domain motions in various NOS proteins. Furthermore, we will investigate local conformational adjustments in the docked state. We will then apply our integrated approach to study remodeling of the conformational landscape by functionally important phosphorylation. Taken together, these results will provide a comprehensive quantitative understanding of protein dynamics as a central part of NOS mechanisms. The proposed research is significant as it will answer long-standing fundamental questions about the NOS isoforms by defining the conformational aspects (statistics, dynamics, and energy landscape) that govern the obligatory electron transfer steps in NOS. This work will positively impact our understanding of other biomolecules as defining structure-dynamics-function relationship lies at the heart of current biochemical research.

项目总结

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Interdomain Interactions Modulate the Active Site Dynamics of Human Inducible Nitric Oxide Synthase.

DOI：
10.1021/acs.jpcb.2c04091
发表时间：
2022-09-15
期刊：
JOURNAL OF PHYSICAL CHEMISTRY B
影响因子：
3.3
作者：
Tumbic, Goran W.;Li, Jinghui;Jiang, Ting;Hossan, Md Yeathad;Feng, Changjian;Thielges, Megan C.
通讯作者：
Thielges, Megan C.

An isoform-specific pivot modulates the electron transfer between the flavin mononucleotide and heme centers in inducible nitric oxide synthase.

DOI：
10.1007/s00775-020-01824-w
发表时间：
2020-12
期刊：
Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry
影响因子：
0
作者：
Zheng H;Li J;Feng C
通讯作者：
Feng C

Probing Protein Dynamics in Neuronal Nitric Oxide Synthase by Quantitative Cross-Linking Mass Spectrometry.

DOI：
10.1021/acs.biochem.3c00245
发表时间：
2023-08-01
期刊：
BIOCHEMISTRY
影响因子：
2.9
作者：
Jiang, Ting;Wan, Guanghua;Zhang, Haikun;Gyawali, Yadav Prasad;Underbakke, Eric S.;Feng, Changjian
通讯作者：
Feng, Changjian

Heat shock protein 90α increases superoxide generation from neuronal nitric oxide synthases.