Structural Basis of Dilated Cardiomyopathy

扩张型心肌病的结构基础

• 批准号: 10183307
• 负责人: Gianluigi Veglia
• 金额: $ 38.24万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10183307
• 关键词: ATP phosphohydrolase; Acylation; Adrenergic Agents; Affect; Affinity; Binding; Biochemical; Biological Assay; Biophysics; Ca(2+)-Transporting ATPase; Calorimetry; Cardiac; Cardiac Myocytes; Cardiac Output; Cardiotoxicity; Cause of Death; Cells; Cellular biology; Collaborations; Complex; Cyclic AMP-Dependent Protein Kinases; Cytoplasmic Tail; DNA Sequence Alteration; Diagnosis; Diastole; Dilated Cardiomyopathy; Equilibrium; Fluorescence; Fluorescence Resonance Energy Transfer; Functional disorder; Genetic Predisposition to Disease; Goals; Heart; Heart Contractilities; Heart failure; Homeostasis; Hot Spot; Human; Human Genetics; In Vitro; Knowledge; Lead; Left; Link; Magic; Maps; Membrane; Membrane Proteins; Methods; Modeling; Modification; Molecular; Molecular Biology; Molecular Structure; Morbidity - disease rate; Mutation; Myocardium; NMR Spectroscopy; Nuclear; Oligonucleotides; Pathogenicity; Pathologic; Patients; Phenotype; Phosphorylation; Physiological; Positioning Attribute; Post-Translational Protein Processing; RNA; Regulation; Relaxation; Research Proposals; Resolution; Sarcoplasmic Reticulum; Signal Pathway; Signal Transduction; Structure; Testing; Therapeutic; Therapeutic Intervention; Titrations; Translating; Vertebral column; atomic interactions; base; design; diabetic; diabetic patient; experimental study; familial dilated cardiomyopathy; genetic regulatory protein; glycosylation; heart function; in vivo; inorganic phosphate; intermolecular interaction; mortality; mutant; novel therapeutic intervention; phospholamban; response; small molecule; solid state nuclear magnetic resonance

ABSTRACT The overall goal of this research proposal is to determine the molecular and structural determinants for dilated cardiomyopathy (DCM) linked to phospholamban (PLN), a membrane protein involved in Ca2+ transport in the sarcoplasmic reticulum (SR). PLN binds and reversibly inhibits the SR Ca-ATPase (SERCA), regulating heart diastole in response to Ca2+ and -adrenergic signaling pathways. -adrenergic control of PLN via reversible phosphorylation at Ser16 relieves PLN’s inhibitory effects to restore SERCA’s basal activity, while shifts in cytosolic Ca2+ levels modulate PLN interactions with SERCA. PLN’s position at the crossroads of Ca2+ signaling and -adrenergic stimulation makes it a key regulator for cardiac output. Here, we propose to analyze the effects of new lethal mutants of PLN (PLNR9H, PLNR9L, and PLNR25C; AIM1) as well as its post-translational modifications (O-linked N-Glycosylation at Ser16 and S-Acylation at Cys36; AIM2) found in patients diagnosed with DCM and understand how they affect Ca2+ cycling. We will employ an integrated approach including biochemical, molecular biology, and spectroscopic methods (FRET, solution NMR, magic angle spinning, and oriented solid-state NMR) in concert with both in cell and in vivo experiments carried out independently by our collaborators. These experiments will determine the cardiotoxic mechanisms of these new mutants and PTMs to obtain a new, unifying regulatory model for cardiac contractility, bridging the Ca2+ and -adrenergic signaling pathways. Finally, in AIM3, we will investigate how to reverse PLN inhibito- ry effects as a mean to augment cardiac contractility under pathological conditions. The scientific premise of the latter AIM is based on our recent discovery that single-stranded oligonucleotides modulate PLN’s regula- tion of SERCA. Together, these studies will pave the way for understanding how mutations and PTMs disrupt Ca2+ and/or - adrenergic signaling, and how cardiac function may be rescued by restoring Ca2+ homeostatic balance.

摘要

项目成果

PHRONESIS: A One-Shot Approach for Sequential Assignment of Protein Resonances by Ultrafast MAS Solid-State NMR Spectroscopy.

• DOI: 10.1002/cphc.202200127
• 发表时间: 2022-07-05
• 期刊: Chemphyschem : a European journal of chemical physics and physical chemistry

T2* weighted Deconvolution of NMR Spectra: Application to 2D Homonuclear MAS Solid-State NMR of Membrane Proteins.

NMR 谱图的 T2* 加权解卷积：在膜蛋白的 2D 同核 MAS 固态 NMR 中的应用。

• DOI: 10.1038/s41598-019-44461-3
• 发表时间: 2019
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: VS,Manu;Gopinath,Tata;Wang,Songlin;Veglia,Gianluigi
• 通讯作者: Veglia,Gianluigi

Cytoplasmic nucleic acid-based XNAs directly enhance live cardiac cell function by a Ca2+ cycling-independent mechanism via the sarcomere.

基于细胞质核酸的 XNA 通过肌节的 Ca2 循环独立机制直接增强活心脏细胞功能。

• DOI: 10.1016/j.yjmcc.2019.02.016
• 发表时间: 2019
• 期刊: Journal of molecular and cellular cardiology
• 影响因子: 5
• 作者: Thompson,BrianR;Soller,KaileyJ;Vetter,Anthony;Yang,Jing;Veglia,Gianluigi;Bowser,MichaelT;Metzger,JosephM
• 通讯作者: Metzger,JosephM

Proton-detected polarization optimized experiments (POE) using ultrafast magic angle spinning solid-state NMR: Multi-acquisition of membrane protein spectra.

使用超快魔角旋转固态 NMR 的质子检测偏振优化实验 (POE)：膜蛋白光谱的多次采集。

• DOI: 10.1016/j.jmr.2019.106664
• 发表时间: 2020
• 期刊: Journal of magnetic resonance (San Diego, Calif. : 1997)
• 作者: Gopinath,T;Veglia,Gianluigi
• 通讯作者: Veglia,Gianluigi

CHESPA/CHESCA-SPARKY: automated NMR data analysis plugins for SPARKY to map protein allostery

CHESPA/CHESCA-SPARKY：SPARKY 的自动化 NMR 数据分析插件，用于绘制蛋白质变构图

• DOI: 10.1093/bioinformatics/btaa781
• 发表时间: 2020
• 期刊: Bioinformatics
• 影响因子: 5.8
• 作者: Shao, Hongzhao;Boulton, Stephen;Olivieri, Cristina;Mohamed, Hebatallah;Akimoto, Madoka;Subrahmanian, Manu Veliparambil;Veglia, Gianluigi;Markley, John L;Melacini, Giuseppe;Lee, Woonghee
• 通讯作者: Lee, Woonghee