The applicant proposes a two aim strategy to explore molecular mechanisms of the regulation of ion-motive ATPases.
Aim 1 focuses on the cardiac calcium pump (SERCA) and its regulation by phospholamban (PLB). The proposed research will use fluorescence resonance energy transfer (FRET) and chemical crosslinking to test the hypothesis that PLB binds to multiple sites on SERCA, and that these discrete binding sites mediate different functional effects.
Aim 2 describes a novel high throughput assay for investigation of the binding interactions of phospholemman (PLM) with itself and its regulatory target, the sodium/potassium pump (NKA). The goal of the proposed experiments are to develop a novel peptide superinhibitor of NKA as a safer alternative to cardiac glycosides, a class of inotropic drugs used in the treatment of heart failure. The experiments described in the two Aims of this application will provide new insight into fundamental mechanisms of regulation of ion-motive ATPases, and may improve our understanding of the ion transport disorders associated with heart failure.

Public Health Relevance

This research project focuses on two enzymes that are critical for the function of the human heart. Both of these related enzymes are ion pumps;one transports calcium, and the other transports sodium. Ion pumps are of great clinical significance because of their central role in cardiac function and disease. Disorders of these proteins are associated with heart failure, a leading cause of human mortality, so they are considered high value therapeutic targets. The proposed experiments will reveal new information about the mechanisms that regulate the function of ion pumps. The knowledge gained in this research may yield new intervention strategies that will improve human health.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01HL092321-07
Application #
8677619
Study Section
Biochemistry and Biophysics of Membranes Study Section (BBM)
Program Officer
Krull, Holly
Project Start
Project End
Budget Start
Budget End
Support Year
7
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Loyola University Chicago
Department
Physiology
Type
Schools of Medicine
DUNS #
City
Maywood
State
IL
Country
United States
Zip Code
60153
Blackwell, Daniel J; Zak, Taylor J; Robia, Seth L (2016) Cardiac Calcium ATPase Dimerization Measured by Cross-Linking and Fluorescence Energy Transfer. Biophys J 111:1192-202
Himes, Ryan D; Smolin, Nikolai; Kukol, Andreas et al. (2016) L30A Mutation of Phospholemman Mimics Effects of Cardiac Glycosides in Isolated Cardiomyocytes. Biochemistry 55:6196-6204
Dvornikov, Alexey V; Smolin, Nikolai; Zhang, Mengjie et al. (2016) Restrictive Cardiomyopathy Troponin I R145W Mutation Does Not Perturb Myofilament Length-dependent Activation in Human Cardiac Sarcomeres. J Biol Chem 291:21817-21828
Abrol, Neha; de Tombe, Pieter P; Robia, Seth L (2015) Acute inotropic and lusitropic effects of cardiomyopathic R9C mutation of phospholamban. J Biol Chem 290:7130-40
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
Iram, Surtaj H; Gruber, Simon J; Raguimova, Olga N et al. (2015) ATP-Binding Cassette Transporter Structure Changes Detected by Intramolecular Fluorescence Energy Transfer for High-Throughput Screening. Mol Pharmacol 88:84-94
Abrol, Neha; Smolin, Nikolai; Armanious, Gareth et al. (2014) Phospholamban C-terminal residues are critical determinants of the structure and function of the calcium ATPase regulatory complex. J Biol Chem 289:25855-66
Bossuyt, Julie; Chang, Chia-Wei; Helmstadter, Kathryn et al. (2011) Spatiotemporally distinct protein kinase D activation in adult cardiomyocytes in response to phenylephrine and endothelin. J Biol Chem 286:33390-400
Xu, Li; Pallikkuth, Sandeep; Hou, Zhanjia et al. (2011) Dysferlin forms a dimer mediated by the C2 domains and the transmembrane domain in vitro and in living cells. PLoS One 6:e27884
Ha, Kim N; Masterson, Larry R; Hou, Zhanjia et al. (2011) Lethal Arg9Cys phospholamban mutation hinders Ca2+-ATPase regulation and phosphorylation by protein kinase A. Proc Natl Acad Sci U S A 108:2735-40

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