Proton NMR and CD solution conformation determination and opioid receptor binding studies of a dynorphin A(1-17) model peptide.
The opioid peptide dynorphin A(1-17) contains a peptide segment in residues 7-15 with the potential to form an amphiphilic beta-strand. This amphiphilic structure may, like the amphiphilic alpha-helices found in many other peptide hormones, be an important determinant of its interactions with membranes and receptors. In order to investigate and characterize these interactions, we have synthesized a 17-residue dynorphin analogue (YGGFLKKVKPKVKVKSS) that incorporates a peptide model of this amphiphilic secondary structure with minimized homology (25%) relative to the native sequence. This peptide exhibits the full biological potency of dynorphin in assays of kappa-opioid receptor binding, and is more selective for this type of opioid receptor than the natural peptide. The conformation of the model peptide in aqueous solution has been investigated in detail by NMR spectroscopy. The values of the NH-CH alpha coupling constants together with rotating frame NOEs indicate the presence of an amphiphilic structure together with some beta-strand structure in residues 7-15, and demonstrate that a peptide model that stabilizes this structure in aqueous solution and enhances kappa-opioid receptor selectivity can be successfully designed using using alternating lysine and valine residues.
Vaughn
J B
JB
Laboratory of Bioorganic Chemistry and Biochemistry, Rockefeller University, New York, NY 10021.
Taylor
J W
JW
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002942:Circular Dichroism; D004399:Dynorphins / Q000378:metabolism*; D006168:Guinea Pigs; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis; D011487:Protein Conformation; D011957:Receptors, Opioid / Q000378:metabolism*; D001615:beta-Endorphin / Q000378:metabolism
D010455:Peptides; D011957:Receptors, Opioid; D001615:beta-Endorphin; D004399:Dynorphins