Effects of naturally occurring arginine 14 deletion on phospholamban conformational dynamics and membrane interactions

Vitaly V. Vostrikov, Kailey J. Soller, Kim N. Ha, T. Gopinath, Gianluigi Veglia

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

Phospholamban (PLN) is a single-pass membrane protein that regulates the sarco(endo)plasmic reticulum Ca2 +-ATPase (SERCA). Phosphorylation of PLN at Ser16 reverses its inhibitory function under β-adrenergic stimulation, augmenting Ca2 + uptake in the sarcoplasmic reticulum and muscle contractility. PLN exists in two conformations; a T state, where the cytoplasmic domain is helical and adsorbed on the membrane surface, and an R state, where the cytoplasmic domain is unfolded and membrane detached. Previous studies have shown that the PLN conformational equilibrium is crucial to SERCA regulation. Here, we used a combination of solution and solid-state NMR to compare the structural topology and conformational dynamics of monomeric PLN (PLNAFA) with that of the PLNR14del, a naturally occurring deletion mutant that is linked to the progression of dilated cardiomyopathy. We found that the behavior of the inhibitory transmembrane domain of PLNR14del is similar to that of the native sequence. Conversely, the conformational dynamics of R14del both in micelles and lipid membranes are enhanced. We conclude that the deletion of Arg14 in the cytoplasmic region weakens the interactions with the membrane and shifts the conformational equilibrium of PLN toward the disordered R state. This conformational transition is correlated with the loss-of-function character of this mutant and is corroborated by SERCA's activity assays. These findings support our hypothesis that SERCA function is fine-tuned by PLN conformational dynamics and begin to explain the aberrant regulation of SERCA by the R14del mutant. This article is part of a Special Issue entitled: NMR Spectroscopy for Atomistic Views of Biomembranes and Cell Surfaces. Guest Editors: Lynette Cegelski and David P. Weliky.

Original languageEnglish (US)
Pages (from-to)315-322
Number of pages8
JournalBiochimica et Biophysica Acta - Biomembranes
Volume1848
Issue number1
DOIs
StatePublished - Jan 2015

Bibliographical note

Funding Information:
This work is supported by a grant from the National Institute of Health ( GM072701 ) and fellowships from the American Heart Association ( 13POST14670054 to V.V. and 13PRE16950023 to K.S.) and the National Heart Lung and Blood Institute ( 5F31HL095361 to K.H.). The NMR spectra were acquired at the Minnesota NMR Center. G.V., V.V., K.S., K.H. and T.G. designed and performed the experiments. V.V. and K.S. analyzed the data. V.V. and G.V. wrote the manuscript with input from K.S., K.H., and T.G.

Publisher Copyright:
© 2014 Elsevier B.V. All rights reserved.

Keywords

  • Dilated cardiomyopathy
  • Membrane proteins
  • Phospholamban
  • R14 deletion
  • SERCA
  • Solid-state NMR

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