Binding of peptide fragments from a seven helix membrane receptor to lipid bilayers and to micelles.
Membrane proteins influence the organizational and motional properties of lipids, while the conformation and function of these proteins (receptors, channels, enzymes, pumps) are affected by the lipid environment. Model systems consisting of peptides and lipids can provide information at a molecular level about the interactions between proteins and lipids in biological membranes. We have synthesized peptides (residues 253-266 (EYWSTFGNLHHISL) from the seven-helix receptor expressed by the mas oncogene), having free or blocked N- and C-terminals. An analog was obtained by linking a spin-labeled amino acid to the N-terminal via a peptide bond. Several spectroscopic techniques were employed to study the interaction between the peptides and lipophilic systems (zwitterionic and negatively charged phospholipid bilayers, and negatively charged, positively charged, zwitterionic and nonionic micelles). Peptide conformational changes were monitored by circular dichroism (CD). The peptides acquired an increased secondary structure upon binding to the lipid systems. Additional evidence for peptide incorporation into micelles came from fluorescence measurements which indicated a blue shift of the tryptophan's emission wavelength, and from ESR spectra of the spin-labeled analog. While narrow lines were obtained in the aqueous phase, line broadening indicative of slower motion was observed in the presence of the lipophilic aggregates. The slow exchange between the two media allowed the evaluation of partition coefficients. The spectra in aqueous solution were also sensitive to conformational changes as a function of pH, allowing the determination of the N-terminal pK. ESR spectra of lipid spin probes incorporated into phospholipid bilayers indicated that the lipids became more immobilized upon binding of the peptides.
Pertinhez-Sini
T A
TA
Departamento de Bioquímica, Universidade de São Paulo, Brasil.
Nakaie
C R
CR
Carvalho
R S
RS
Paiva
A C
AC
Tabak
M
M
Toma
F
F
Schreier
S
S
D000595:Amino Acid Sequence; D004578:Electron Spin Resonance Spectroscopy; D008051:Lipid Bilayers / Q000378:metabolism*; D008823:Micelles; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism*; D017433:Protein Structure, Secondary; D018000:Receptors, Peptide / Q000737:chemistry* / Q000235:genetics
D008051:Lipid Bilayers; D008823:Micelles; D010446:Peptide Fragments; D018000:Receptors, Peptide