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T Mavromoustakos

Publications and source records attributed to T Mavromoustakos.

63 records · Page 4Linked to original sources
PubMed1993

Topography of tetrahydrocannabinol in model membranes using neutron diffraction.

Small-angle neutron scattering was used to determine the intralamellar location of (-)-delta 8-tetrahydrocannabinol (delta 8-THC) in hydrated dipalmitoylphosphatidylcholine (DPPC) bilayers. Nuclear scattering density profiles were calculated from measurements on deuterium and non-deuterium-labelled inclusions (8.3% (w/w)) of delta 8-THC in DPPC multilayer samples. By comparing pairs of such nuclear density profiles, the locations of the deuterium labels were determined. Present results on the topography of delta 8-THC in membranes are compared with earlier X-ray measurements using iodine labelling. Whereas the position of the phenolic hydroxy group is similar in both types of measurement, a difference is found in the conformation of the terminal methyl groups of the cannabinoid side-chains. The X-ray measurements on dimyristoylphosphatidylcholine (DMPC) indicated that the iodine-labelled cannabinoid side-chains assume an all-trans orientation with the terminal iodine atom pointing inward into the membrane away from the tricyclic region while the neutron measurements indicate that the terminal CH3 group of delta 8-THC aligns itself at the level of the tricyclic ring system implying that the side chain exists in a more compact conformation perpendicular to the DPPC hydrocarbons. A Gaussian function analysis of the data indicates that the delta 8-THC molecule is significantly delocalized in the DPPC membrane in the liquid crystal phase. The mean location of delta 8-THC suggests that the active site on a membrane-embedded receptor protein will lie near the polar interface at the base of the phospholipid headgroups.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1993

Small angle X-ray diffraction studies of (-)-delta 8-tetrahydrocannabinol and its O-methyl analog in membranes.

Small angle X-ray diffraction was used to study the topography of (-)-delta 8-tetrahydrocannabinol (delta 8-THC) and its pharmacologically inactive methoxy analog (-)-O-methyl-delta 8-tetrahydrocannabinol (Me-delta 8-THC) in a membrane. The membrane preparations were partially hydrated dimyristoylphosphatidylcholine (DMPC) bilayers. Comparisons between electron density profiles from the drug-containing and drug-free membranes showed that the amphipathic delta 8-THC residues near the interface of the bilayer where the polar phenolic OH of the drug molecule is oriented towards the corresponding polar side of the phospholipid bilayer. Conversely, the highly hydrophobic Me-delta 8-THC distributes deeper in the bilayer away from the interface. Our results point out these two structurally-related, but pharmacologically very different, cannabinoids interact with membranes in strikingly different manners. This observation may, in part, explain the different pharmacological properties of the two cannabinoids.

Dronabinol↗
PubMed1992

Amphipathic interactions of cannabinoids with membranes. A comparison between delta 8-THC and its O-methyl analog using differential scanning calorimetry, X-ray diffraction and solid state 2H-NMR.

The effects of (-)-delta 8-tetrahydrocannabinol (delta 8-THC) and its biologically inactive O-methyl ether analog on model phospholipid membranes were studied using a combination of differential scanning calorimetry (DSC), small angle X-ray diffraction and solid state 2H-NMR. The focus of this work is on the amphipathic interactions of cannabinoids with membranes and the role of the free phenolic hydroxyl group which is the only structural difference between these two cannabinoids. Identically prepared aqueous multilamellar dispersions of phosphatidylcholines in the absence and presence of cannabinoids were used. The DSC thermograms and X-ray diffraction patterns of these preparations allowed us to detect the strikingly different manners in which these two cannabinoids affect the thermotropic properties and the thickness of the bilayer. In order study the effects of the cannabinoids on different regions of the bilayer, we used solid state 2H-NMR with four sets of model membranes from dipalmitoylphosphatidylcholine deuterated in different sites, viz., the choline trimethylammonium head group, or one of the following three groups in the acyl chains; the 2'-methylene, 7'-methylene, 16'-methyl groups. Analysis of quadrupolar splittings indicated that delta 8-THC resides near the bilayer interface and the inactive analog sinks deeper towards the hydrophobic region. The temperature dependence of the solid state 2H-NMR spectra showed that, during the bilayer phase transition, the disordering of the choline head groups is a separate event from the melting of the acyl chains, and that amphipathic interactions between delta 8-THC and the membrane separate these two events further apart in temperature. The inactive analog lacks the ability to induce such a perturbation.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1992

Bilayer structure and physical dynamics of the cytochrome b5 dimyristoylphosphatidylcholine interaction.

Cytochrome b5 is a microsomal membrane protein which provides reducing potential to delta 5-, delta 6-, and delta 9-fatty acid desaturases through its interaction with cytochrome b5 reductase. Low angle x-ray diffraction has been used to determine the structure of an asymmetrically reconstituted cytochrome b5:DMPC model membrane system. Differential scanning calorimetry and fluorescence anisotropy studies were performed to examine the bilayer physical dynamics of this reconstituted system. These latter studies allow us to constrain structural models to those which are consistent with physical dynamics data. Additionally, because the nonpolar peptide secondary structure remains unclear, we tested the sensitivity of our model to different nonpolar peptide domain configurations. In this modeling approach, the nonpolar peptide moiety was arranged in the membrane to meet such chemically determined criteria as protease susceptibility of carboxyl- and amino-termini, tyrosine availability for pH titration and tryptophan 109 location, et cetera. In these studies, we have obtained a reconstituted cytochrome b5:DMPC bilayer structure at approximately 6.3 A resolution and conclude that the nonpolar peptide does not penetrate beyond the bilayer midplane. Structural correlations with calorimetry, fluorescence anisotropy and acyl chain packing data suggest that asymmetric cytochrome b5 incorporation into the bilayer increases acyl chain order. Additionally, we suggest that the heme peptide:bilayer interaction facilitates a discreet heme peptide orientation which would be dependent upon phospholipid headgroup composition.

Biophysical Phenomena↗
PubMed1991

Small angle x-ray diffraction studies on the topography of cannabinoids in synaptic plasma membranes.

In a previous publication, we have described in detail how we used small angle x-ray diffraction to determine the topography of (-)-delta 8-tetrahydrocannabinol (delta 8-THC) in dimyristoylphosphatidylcholine (DMPC) bilayers, and to deduce the conformation of the THC side chain by using the iodo-analog (5'-I-delta 8-THC) in the model membrane. We have now extended our studies to synaptic plasma membrane systems where the cannabinoids are believed to exert part of their pharmacological effects. Synaptic plasma membranes (SPM) were isolated from fresh bovine brains and delta 8-THC was incorporated into the membranes. By comparing the electron density profiles of drug free and drug-containing SPM preparations, we observed an electron density increase due to the presence of delta 8-THC in a region centered at 9.2 A from the terminal methyl groups of the membrane bilayer. In an attempt to dissect the effects of different membrane components on the topography of delta 8-THC, we carried out parallel experiments using membrane preparations from the synaptosomal membrane total lipid extract (TLX) as well as from bovine brain phosphatidyl choline extract (PCX) containing 30 mole percent cholesterol (Chol). Our results regarding the topography of delta 8-THC and 5'-I-delta 8-THC in these lipid membranes show that the TLX bilayer simulates the natural membrane environment very closely whereas in the PCX/Chol bilayer delta 8-THC resides at a location approximately 4 A closer to the membrane interface, similar to that found in our previous study using DMPC model membrane.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

Study of the topography of cannabinoids in model membranes using X-ray diffraction.

Small-angle X-ray diffraction was used to determine the topography of (-)-delta 8-tetrahydrocannabinol in partially hydrated dimyristoylphosphatidylcholine bilayers. Electron density profiles of lipid bilayers in the presence and absence of the cannabinoid were calculated using Fourier transform. Step-function equivalent profiles were then constructed to obtain the absolute electron density scale. We have compared the electron density profiles of the above preparations to determine the location of the drug molecule in the bilayer. By using (-)-5'-iodo-delta 8-tetrahydrocannabinol in parallel experiments, we were also able to locate the iodine atom in the bilayer and deduce the conformation of the cannabinoid side alkyl chain. All comparisons were made between different preparations having the same mesomorphic form and total period repeat distance. To achieve this, we have carried out X-ray diffraction experiments at various temperatures to cover the different mesomorphic phases and combined our data with the corresponding results from differential scanning calorimetry. Based on the results of this work and previous data on the orientation of the cannabinoid in model membranes, we concluded that the phenolic hydroxy group of the drug molecule exists near the carbonyl groups of DMPC and that the average position of the iodine atom is approx. 5.5 A from the center (terminal methyl region) of the DMPC bilayer. This requires the cannabinoid side-chain to assume an orientation parallel to the bilayer chains.

Calorimetry, Differential Scanning↗
PubMed1990

The molecular features of membrane perturbation by anaesthetic steroids: a study using differential scanning calorimetry, small angle X-ray diffraction and solid state 2H NMR.

We have studied the interactions of the anaesthetic steroid alphaxalone and its inactive isomer delta 16-alphaxalone with model membrane bilayers using differential scanning calorimetry, small angle X-ray diffraction and solid state NMR. Our data show that the anaesthetic steroid broadens the membrane phase transition and increases the ratio of gauche to trans conformers in the membrane. Delta 16-Alphaxalone has only small effects on membrane and incorporates to a limited degree in the bilayer. The amphipathic anaesthetic steroid alphaxalone is located near the membrane interface (the junction of the polar and hydrophobic regions of the phospholipids forming the bilayer). It orients with its long axis parallel to the chains of the lipid membranes and its 3 alpha-hydroxyl group near the sn-2 carbonyl. Anchoring of the steroid at the membrane interface and imperfect packing with the bilayer chains may be involved in membrane perturbation and eventually lead to anaesthesia.

Anesthetics↗
PubMed1988

The conformation of (-) 8 alpha and (-) 8 beta-hydroxy-delta 9-tetrahydrocannabinols and their interactions with model membranes.

8 alpha- and 8 beta-Hydroxy-delta 9-tetrahydrocannabinols (THC's), two metabolites of the naturally occurring delta 9-THC have been shown to possess differences in pharmacological activity. We have studied the conformations of these two compounds, as well as their interactions with model membrane systems and compared them with delta 9-THC. The conformational study, carried out in solution and using high resolution NMR indicated that differences in the ring conformations of these two compounds were negligible but that the 8-hydroxy group of the 8 beta-OH compound extended approximately 1.4A higher above the plane of the aromatic ring than in the 8 alpha-OH isomer. This difference could prove significant in the interaction of these molecules with lipid bilayers. We found that both 8 alpha- and 8 beta-OH analogs affected the melting behavior of hydrated DPPC bilayers including a lowering of the main transition temperature (Tc), a broadening of that transition and the abolishment of the pretransition of DPPC. The effects of the more active compound, 8 beta-OH-delta 9-THC on the model membrane approximated closely those of delta 9-THC, while the less active 8 alpha-OH epimer produced different thermotropic changes.

1,2-Dipalmitoylphosphatidylcholine↗