Structure and topology of a peptide segment of the 6th transmembrane domain of the Saccharomyces cerevisae α-factor receptor in phospholipid bilayers

Kathleen G. Valentine, Shi Feng Liu, Francesca M. Marassi, Gianluigi Veglia, Stanley J. Opella, Fa Xiang Ding, Shu Hua Wang, Boris Arshava, Jeffrey M. Becker, Fred Naider

Research output: Contribution to journalArticlepeer-review

39 Scopus citations

Abstract

A detailed analysis of the structure of an 18-residue peptide AQSLLVPSIIFILAYSLK [M6(252-269, C252A)] in 1,2-dimyristoyl-sn-glycero-phosphocholine bilayers was carried out using solid state NMR and attenuated total reflection Fourier transform infrared spectroscopy. The peptide corresponds to a portion of the 6th transmembrane domain of the α-factor receptor of Saccharomyces cerevisiae. Ten homologs of M6(252-269, C252A) were synthesized in which individual residues were labeled with 15N. One- and two-dimensional solid state NMR experiments were used to determine the chemical shifts and 1H-15N dipolar coupling constants for the 15N-labeled peptides in oriented dimyristoylphosphatidylcholine bilayers on stacked glass plates. These parameters were used to calculate the structure and orientation of M6(252-269, C252A) in the bilayers. The results indicate that the carboxyl terminal residues (9-14) are α-helical and oriented with an angle of about 8° with respect to the bilayer normal. Independently, an attenuated total reflection Fourier transform infrared spectroscopy analysis on M6(252-269, C252A) in a 1,2-dimyristoyl-sn-glycero-phosphocholine bilayer concluded that the helix tilt angle was about 12.5°. The results on the structure of M6(252-269, C252A) in bilayers are in good agreement with the structure determined in trifluoroethanol/water solutions (B. Arshava et al. Biopolymers, 1998, Vol. 46, pp. 343-357). The present study shows that solid state NMR spectroscopy can provide high resolution information on the structure of transmembrane domains of a G protein-coupled receptor.

Original languageEnglish (US)
Pages (from-to)243-256
Number of pages14
JournalBiopolymers
Volume59
Issue number4
DOIs
StatePublished - Oct 5 2001

Keywords

  • 6th transmembrane peptide
  • Phospholipid bilayers, ATR-FTIR
  • Saccharomyces cerevisae
  • Solid state NMR
  • α-Factor receptor

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