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Structure and topology of a peptide segment of the 6th transmembrane domain of the Saccharomyces cerevisae α‐factor receptor in phospholipid bilayers
Author(s) -
Valentine Kathleen G.,
Liu ShiFeng,
Marassi Francesca M.,
Veglia Gianluigi,
Opella Stanley J.,
Ding FaXiang,
Wang ShuHua,
Arshava Boris,
Becker Jeffrey M.,
Naider Fred
Publication year - 2001
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/1097-0282(20011005)59:4<243::aid-bip1021>3.0.co;2-h
Subject(s) - chemistry , lipid bilayer , bilayer , transmembrane protein , crystallography , transmembrane domain , solid state nuclear magnetic resonance , nuclear magnetic resonance spectroscopy , phospholipid , peptide , attenuated total reflection , spectroscopy , infrared spectroscopy , membrane , analytical chemistry (journal) , stereochemistry , nuclear magnetic resonance , receptor , chromatography , organic chemistry , biochemistry , physics , quantum mechanics
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 15 N. One‐ and two‐dimensional solid state NMR experiments were used to determine the chemical shifts and 1 H– 15 N dipolar coupling constants for the 15 N‐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. © 2001 John Wiley & Sons, Inc. Biopolymers 59: 243–256, 2001