Determination of helical membrane protein topology using residual dipolar couplings and exhaustive search algorithm: Application to phospholamban

Alessandro Mascioni, Becky L. Eggimann, Gianluigi Veglia

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

Dipolar waves are distinct hallmarks of both the secondary and tertiary structures of α-helical proteins that are immobilized in membrane bilayers or embedded in anisotropic media. We present a simple, semi-empirical approach that exploits the modulation of the amplitude and average of dipolar waves to determine the topology of α-helical proteins. Moreover, we describe the application of this method for the structural determination of a detergent solubilized membrane protein, phospholamban (PLB) that is involved in calcium regulation of cardiac muscle. When combined with high-resolution solid-state NMR data, this method can serve as a fast route for determining the topology of helical membrane proteins solubilized in detergent micelles.

Original languageEnglish (US)
Pages (from-to)133-144
Number of pages12
JournalChemistry and Physics of Lipids
Volume132
Issue number1
DOIs
StatePublished - Oct 2004

Bibliographical note

Funding Information:
This work was supported in part by grants to G.V. (NIH GM64742; AHA 0160465Z). The authors would like to thank Dr. S.J. Opella, Francesca Marassi, Michael Mesleh for many helpful discussions, Roberto Di Fonzo for encouragement and support, David Live and Beverly Ostrowski for help with NMR experiments. We also benefited from the facilities at the Minnesota High-Field NMR Center in the Department of Biochemistry, Molecular Biology, and Biophysics; University of Minnesota. NMR instrumentation was provided with funds from the NSF (BIR-961477) and the University of Minnesota Medical School.

Keywords

  • Ca-ATPase
  • DPC micelles
  • Dipolar waves
  • Exhaustive search
  • Helical wheels
  • Membrane proteins
  • NMR
  • Phospholamban
  • Polyacrylamide gel
  • Residual dipolar couplings
  • SERCA
  • α-Helix

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