1. Effects ofN- andC-terminal addition of oligolysines or native loop residues on the biophysical properties of transmembrane domain peptides from a G-protein coupled receptor
- Author
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V. V. Sureshbabu, Patricia Cano-Sanchez, Boris Arshava, Tatsuya Inui, Beatrice Severino, Joe Russo, Fred Naider, Jeff Becker, Fa-Xiang Ding, P., CANO SANCHEZ, Severino, Beatrice, V. V., Sureshbabu, J., Russo, T., Inui, F. X., Ding, B., Arshava, J., Becker, and F., Naider
- Subjects
transmembrane peptide ,Circular dichroism ,Receptors, Peptide ,Stereochemistry ,Molecular Sequence Data ,Lysine ,Peptide ,Saccharomyces cerevisiae ,Biochemistry ,Micelle ,oligolysines ,Receptors, G-Protein-Coupled ,chemistry.chemical_compound ,Structural Biology ,Spectroscopy, Fourier Transform Infrared ,Drug Discovery ,Polylysine ,Amino Acid Sequence ,Sodium dodecyl sulfate ,Molecular Biology ,Chromatography, High Pressure Liquid ,Micelles ,G protein-coupled receptor ,Pharmacology ,chemistry.chemical_classification ,Vesicle ,Organic Chemistry ,General Medicine ,Peptide Fragments ,circular dichroism ,Crystallography ,Transmembrane domain ,chemistry ,Solvents ,Molecular Medicine ,Dimyristoylphosphatidylcholine - Abstract
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 -a ndC-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 Lysn (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 &S ons, Ltd.
- Published
- 2006
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