Role of hydrophobic substituents in the interaction of opioid Tyr-Tic dipeptide analogs with dodecylphosphocholine micelles. Molecular partitioning in model membrane systems.
The conformational properties of three Tyr-Tic-NH-R dipeptide analogs [where R = (CH2)2-Ph, (CH2)3-Ph or (CH2)2-cHx; Ph = phenyl; cHx = cyclohexyl and Tic = tetrahydroisoquinoline-3-carboxylic acid] have been investigated in purely aqueous solution and in the presence of fully deuterated dodecylphosphocholine micelles. H-Tyr-Tic-NH-(CH2)2-Ph is an opioid delta-agonist, whereas H-Tyr-Tic-NH-(CH2)3-Ph is a fairly potent delta-antagonist. H-Tyr-Tic-NH-(CH2)2-cHx is a less potent delta-antagonist. 1H-NMR spectra revealed that conformers containing cis and trans configurations of the Tyr-Tic peptide bond were present in all compounds in H2O and the H2O/lipid solvent. Analyses of the NMR data for the compounds in H2O indicate that in all three dipeptides the C-terminal substituent is flexible and the Tyr-side-chain adopts a trans orientation in most of the conformations. This promotes a compact Tyr-Tic structure. NOE patterns observed for the compounds in the micelle solution indicate that Tyr has an even greater tendency to assume a trans side chain configuration in the biphasic-solvent system. This feature was more pronounced in the trans conformers than in the cis conformers. Specific lipid-peptide interactions were indicated by NOESY spectra acquired for micelle samples incorporating 20% (by mass) protonated lipid. According to the obtained NOE data, Tyr and Tic form an aromatic cluster which preferentially inserts into the lipid interior of the micelle for the trans conformers of all three dipeptides and for the cis conformer of H-Tyr-Tic-NH-(CH2)2-Ph. For the cis isomers, partitioning of the C-terminal substituents into the lipid phase exhibited more diverse behaviour. The cis conformers of H-Tyr-Tic-NH-(CH2)3-Ph and H-Tyr-Tic-NH-(CH2)2-cHx preferentially anchor to the micelle via their C-terminal substituent, while the corresponding region in H-Tyr-Tic-NH-(CH2)2-Ph remains flexible and immersed in the aqueous phase. The inconsistent mode of peptide-micelle interaction observed for cis conformers of the three compounds studied is explained in terms of differences in their dipeptide-substituent hydrophobicities. The more apolar the substituent, the greater its tendency to preferentially insert into the lipid core of the micelle. Amide-proton temperature coefficients measured for the three peptides revealed differences amongst the cis and trans isomers. The amide proton in the trans conformer of each compound is highly exposed to the aqueous phase in both solvent systems studied, whereas the cis NH proton of each peptide is only partially exposed. These results demonstrate that a subtle structural modification of an active peptide analog can result in dramatic changes of its biological activity and its mode of partitioning at a membrane surface.
Carpenter
K A
KA
Laboratory of Chemical Biology and Peptide Research, Clinical Research Institute of Montreal, Canada.
Wilkes
B C
BC
Weltrowska
G
G
Schiller
P W
PW
D000577:Amides; D003198:Computer Simulation; D007536:Isomerism; D007546:Isoquinolines / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008566:Membranes; D008823:Micelles; D008956:Models, Chemical; D008968:Molecular Conformation; D009292:Narcotic Antagonists / Q000737:chemistry*; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000737:chemistry; D011522:Protons; D017465:Receptors, Opioid, delta / Q000819:agonists* / Q000037:antagonists & inhibitors*; D044005:Tetrahydroisoquinolines; D017186:Titrimetry; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000737:chemistry
C097291:2-tyrosyl-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid; D000577:Amides; D007546:Isoquinolines; D008823:Micelles; D009292:Narcotic Antagonists; D011522:Protons; D017465:Receptors, Opioid, delta; D044005:Tetrahydroisoquinolines; D010767:Phosphorylcholine; D014443:Tyrosine; C028810:dodecylphosphocholine