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Effects of N ‐ and C ‐terminal addition of oligolysines or native loop residues on the biophysical properties of transmembrane domain peptides from a G‐protein coupled receptor
Author(s) -
CanoSanchez Patricia,
Severino Beatrice,
Sureshbabu V. V.,
Russo Joe,
Inui Tatsuya,
Ding FaXiang,
Arshava Boris,
Becker Jeff,
Naider Fred
Publication year - 2006
Publication title -
journal of peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 66
eISSN - 1099-1387
pISSN - 1075-2617
DOI - 10.1002/psc.816
Subject(s) - chemistry , circular dichroism , peptide , transmembrane protein , transmembrane domain , g protein coupled receptor , sodium dodecyl sulfate , lysine , biophysics , protein secondary structure , vesicle , stereochemistry , membrane , biochemistry , amino acid , receptor , biology
Transmembrane domains (TMDs) of G‐protein coupled receptors (GPCRs) have very low water solubility and often aggregate during purification and biophysical investigations. To circumvent this problem many laboratories add oligolysines to the N ‐ and C ‐termini of peptides that correspond to a TMD. To systematically evaluate the effect of the oligolysines on the biophysical properties of a TMD we synthesized 21 peptides corresponding to either the second (TPIFIINQVSLFLIILHSALYFKY) or sixth (SFHILLIMSSQSLLVPSIIFILAYSLK) TMD of Ste2p, a GPCR from Saccharomyces cerevisiae . Added to the termini of these peptides were either Lys n ( n = 1,2,3) or the corresponding native loop residues. The biophysical properties of the peptides were investigated by circular dichroism (CD) spectroscopy in trifluoroethanol–water mixtures, sodium dodecyl sulfate (SDS) micelles and dimyristoylphosphocholine (DMPC)‐dimyristoylphosphoglycerol (DMPG) vesicles, and by attenuated total reflection Fourier transform infrared (ATR‐FTIR) in DMPC/DMPG multilayers. The results show that the conformation assumed depends on the number of lysine residues and the sequence of the TMD. Identical peptides with native or an equal number of lysine residues exhibited different biophysical properties and structural tendencies. Copyright © 2006 European Peptide Society and John Wiley & Sons, Ltd.