TY - JOUR
T1 - Structure and topology of a peptide segment of the 6th transmembrane domain of the Saccharomyces cerevisae α-factor receptor in phospholipid bilayers
AU - Valentine, Kathleen G.
AU - Liu, Shi Feng
AU - Marassi, Francesca M.
AU - Veglia, Gianluigi
AU - Opella, Stanley J.
AU - Ding, Fa Xiang
AU - Wang, Shu Hua
AU - Arshava, Boris
AU - Becker, Jeffrey M.
AU - Naider, Fred
PY - 2001/10/5
Y1 - 2001/10/5
N2 - 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.
AB - 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.
KW - 6th transmembrane peptide
KW - Phospholipid bilayers, ATR-FTIR
KW - Saccharomyces cerevisae
KW - Solid state NMR
KW - α-Factor receptor
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U2 - 10.1002/1097-0282(20011005)59:4<243::AID-BIP1021>3.0.CO;2-H
DO - 10.1002/1097-0282(20011005)59:4<243::AID-BIP1021>3.0.CO;2-H
M3 - Article
C2 - 11473349
AN - SCOPUS:0035813019
SN - 0006-3525
VL - 59
SP - 243
EP - 256
JO - Biopolymers
JF - Biopolymers
IS - 4
ER -