Defining the Intramembrane Binding Mechanism of Sarcolipin to Calcium ATPase Using Solution NMR Spectroscopy

Jarrod J. Buffy, Bethany A. Buck-Koehntop, Fernando Porcelli, Nathaniel Traaseth, David D. Thomas, Gianluigi Veglia

Research output: Contribution to journalArticle

Abstract

Sarcolipin (SLN) is an integral membrane protein that is expressed in both skeletal and cardiac muscle, where it inhibits SERCA (calcium ATPase) by lowering its apparent Ca2+ affinity in a manner similar to that of its homologue phospholamban (PLN). We use solution NMR to map the structural changes occurring within SLN upon interaction with the regulatory target, SERCA, co-reconstituting the two proteins in dodecylphosphocholine (DPC) detergent micelles, a system that preserves the native structure of SLN and the activity of SERCA, with the goal of comparing these interactions with those of the previously studied PLN-SERCA complex. Our analysis of the structural dynamics of SLN in DPC micelles shows this polypeptide to be partitioned into four subdomains: a short unstructured N terminus (residues 1-6), a short dynamic helix (residues 7-14), a more rigid helix (residues 15-26), and an unstructured C terminus (residues 27-31). Upon addition of SERCA, the different domains behave according to their dynamics, molding onto the surface of the enzyme. Remarkably, each domain of SLN behaves in a manner similar to that of the corresponding domains in PLN, supporting the hypothesis that both SLN and PLN bind SERCA in the same groove and with similar mechanisms.

Original languageEnglish (US)
Pages (from-to)420-429
Number of pages10
JournalJournal of Molecular Biology
Volume358
Issue number2
DOIs
StatePublished - Apr 28 2006

Fingerprint

Calcium-Transporting ATPases
Magnetic Resonance Spectroscopy
Micelles
Sarcoplasmic Reticulum Calcium-Transporting ATPases
Detergents
sarcolipin
Myocardium
Membrane Proteins
Skeletal Muscle
Peptides
phospholamban
Enzymes
Proteins

Keywords

  • Ca-ATPase
  • phospholamban
  • sarcolipin
  • SERCA
  • solution NMR

ASJC Scopus subject areas

  • Virology

Cite this

Defining the Intramembrane Binding Mechanism of Sarcolipin to Calcium ATPase Using Solution NMR Spectroscopy. / Buffy, Jarrod J.; Buck-Koehntop, Bethany A.; Porcelli, Fernando; Traaseth, Nathaniel; Thomas, David D.; Veglia, Gianluigi.

In: Journal of Molecular Biology, Vol. 358, No. 2, 28.04.2006, p. 420-429.

Research output: Contribution to journalArticle

Buffy, Jarrod J. ; Buck-Koehntop, Bethany A. ; Porcelli, Fernando ; Traaseth, Nathaniel ; Thomas, David D. ; Veglia, Gianluigi. / Defining the Intramembrane Binding Mechanism of Sarcolipin to Calcium ATPase Using Solution NMR Spectroscopy. In: Journal of Molecular Biology. 2006 ; Vol. 358, No. 2. pp. 420-429.
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