The overall goal of my research program is to determine the molecular and structural determinants of dilated cardiomyopathy (DCM), the leading cause for heart failure (HF) worldwide. Three major groups of cardiac proteins are involved in DCM: cytoskeletal, nuclear, and sarcomeric. Our group focuses on the study of mutations occurring in phospholamban (PLN), a membrane protein involved in calcium transport in the sarcoplasmic reticulum (SR). PLN binds and inhibits the SR Ca-ATPase (SERCA), regulating heart diastole. PLN phosphorylation at Ser16 by protein kinase A (PKA) reverses the inhibitory effects, constituting the primary response to b-adrenergic stimulation in the heart. Disruptions in the phosphorylation cycle progress to HF. The R9C mutation (PLNR9C) and R14 deletion (PLNR14del), both located in the PLN cytoplasmic domain, prevent PLN phosphorylation and lead to DCM via an unknown mechanism. Using a battery of biochemical, molecular biology, and spectroscopic methods, we will characterize the structural and dynamic effects of these mutations in the formation of the PKA/PLN complex and link them to their cardiotoxicity. Specifically, we plan to decipher the PLN recognition and phosphorylation mechanisms by the kinase in the presence and absence of these deadly mutations. Moreover, we will elucidate the role of lipid membranes and spatial localization via myristoylation in the phosphorylation process. These studies will set the groundwork for understanding the physiology and pathophysiology of phosphorylation signaling in the heart.

Public Health Relevance

Dilated cardiomyopathy (DCM) is the leading cause for heart failure (HF) worldwide. This proposal seeks to elucidate the molecular determinants for DCM using biophysical, biochemical, and molecular biology approaches. Understanding the genesis of the disease at the atomic level will help translate this knowledge into molecular design of new therapeutic approaches to counteract HF.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM072701-05A1
Application #
8288427
Study Section
Macromolecular Structure and Function C Study Section (MSFC)
Program Officer
Chin, Jean
Project Start
2007-06-01
Project End
2016-02-29
Budget Start
2012-05-01
Budget End
2013-02-28
Support Year
5
Fiscal Year
2012
Total Cost
$308,350
Indirect Cost
$99,776
Name
University of Minnesota Twin Cities
Department
Biochemistry
Type
Schools of Medicine
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Kim, Jonggul; Li, Geoffrey; Walters, Michael A et al. (2016) Uncoupling Catalytic and Binding Functions in the Cyclic AMP-Dependent Protein Kinase A. Structure 24:353-63
Soller, Kailey J; Yang, Jing; Veglia, Gianluigi et al. (2016) Reversal of Phospholamban Inhibition of the Sarco(endo)plasmic Reticulum Ca2+-ATPase (SERCA) Using Short, Protein-interacting RNAs and Oligonucleotide Analogs. J Biol Chem 291:21510-21518
Kim, Jonggul; Wang, Yingjie; Li, Geoffrey et al. (2016) A Semiautomated Assignment Protocol for Methyl Group Side Chains in Large Proteins. Methods Enzymol 566:35-57
Gopinath, T; Veglia, Gianluigi (2016) Multiple acquisitions via sequential transfer of orphan spin polarization (MAeSTOSO): How far can we push residual spin polarization in solid-state NMR? J Magn Reson 267:1-8
Soller, Kailey J; Verardi, Raffaello; Jing, Meng et al. (2015) Rheostatic Regulation of the SERCA/Phospholamban Membrane Protein Complex Using Non-Coding RNA and Single-Stranded DNA oligonucleotides. Sci Rep 5:13000
Kim, Jonggul; Masterson, Larry R; Cembran, Alessandro et al. (2015) Dysfunctional conformational dynamics of protein kinase A induced by a lethal mutant of phospholamban hinder phosphorylation. Proc Natl Acad Sci U S A 112:3716-21
Gopinath, T; Mote, Kaustubh R; Veglia, Gianluigi (2015) Simultaneous acquisition of 2D and 3D solid-state NMR experiments for sequential assignment of oriented membrane protein samples. J Biomol NMR 62:53-61
Manu, V S; Veglia, Gianluigi (2015) Genetic algorithm optimized triply compensated pulses in NMR spectroscopy. J Magn Reson 260:136-43
Gopinath, T; Veglia, Gianluigi (2015) Multiple acquisition of magic angle spinning solid-state NMR experiments using one receiver: application to microcrystalline and membrane protein preparations. J Magn Reson 253:143-53
Vostrikov, Vitaly V; Soller, Kailey J; Ha, Kim N et al. (2015) Effects of naturally occurring arginine 14 deletion on phospholamban conformational dynamics and membrane interactions. Biochim Biophys Acta 1848:315-22

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