Fusion and lipid exchange in vesicles containing lipophilic spin labels

Lipophilic non-electrolyte spin labels greatly accelerate the fusion of unilamellar vesicles of dipalmitoylphosphatidylcholine when the system is maintained below the lipid phase transition. Differential scanning calorimetry and centrifugation measurements show that the transformed vesicles are large and probably unilamellar. Differential scanning calorimetry and fluorescence depolarization measurements were also carried out on mixtures of labeled dipalmitoylphosphatidylcholine vesicles and of vesicles composed of pure dimyristoylphosphatidylcholine. A mixing of the membrane components is observed when the vesicles are incubated above the transition temperature of the two constituent lipids. However, the process does not involve a real fusion of the entire vesicles. An exchange of lipid and label monomers between the two lipid phases seems to occur. These observations are discussed in view of the molecular organization of the spin label within the dipalmitoylphosphatidylcholine matrix below and above the lipid transition temperature.     

Structural analogies between protein kinase C activators

Phorbol esters and diacylglycerols activate protein kinase C but specific structural parameters appear to be required for the enzyme activation. We have analyzed the conformation of potent and not potent diacylglycerols and phorbol esters. The orientation of the CH20H group at C3 of 1,2 diolein is remarkably similar to that of the same group at C-20 of 4 beta phorbol didecanoate and crucial for potency in activating the enzyme. Our data suggest that the new conformational approach here described could be used to rationally design specific inhibitors preventing the effects of tumor promoters and to predict the structure of potential tumor promoters.     

The importance of historical myths for the ethnic consciousness of Romanians and ethnic Hungarians in Transylvania

Conclusion The ethnic Hungarians and the Romanians share a number of myths about their ancestors and about their homeland, Transylvania. I fully agree with Anthony D. Smith, who states that the core of ethnic identity is made up of a myth-symbol complex, consisting of myths, symbols, historical memory and key values. According to Smith, myths and symbols guarantee the preservation and the passing down to future generations of ethnic identity.²⁵ The main problem with the ethnic Hungarians and the Romanians of Transylvania is that their convictions are not compatible, they are even contradictory. In the ethnic Hungarians' view, Transylvania is in fact Hungarian, but it was taken from them by Romania in 1920. According to the Romanians, Transylvania is Romanian but for centuries mistakenly considered as Hungarian by the Hungarians. The historical myths are part of the Hungarian as well as the Romanian collective consciousness. They strengthen cohesion within these groups but, as the convictions are incompatible, they widen the gap between them. In my view, these contradictory views are an additional cause of the current mutual oversensitivity, the mutual suspicion and mutual ignorance which characterize the relation between the two ethnic groups. I think that the success of the Romanian nationalist parties such as the PUNR and România Mare ²⁶ among the Romanian villagers of Transylvania ²⁷is partly due to their exploitation of these contradictory myths.However, the differing oral traditions in the villages do not cause open conflicts. That everyone in his own group passes on his own version helps to explain this. In mixed villages like Dumbrava and Mànàstireni, the sensitive issues are not discussed in ethnically mixed groups. Virtually everything to do with history is suppressed in the day-to-day life of the village. Nevertheless, that does not stop one ethnic group from gossiping about the other. Thus, mutual distrust persists. Such a sense of fear is a dangerous breeding-ground because it can easily be exploited by nationalist parties. When anything occurs in politics which appeals to these feelings of fear and distrust, two fronts are lined up in no time. This actually happened as a result of the controversy about the archaeological excavations in Cluj.Fortunately, there is a sufficiently large number of people who realize that this is pointless and who see through the extremist nationalist party leaders' malice. Bearing in mind the tragic example of the former Yugoslavia, they advocate understanding and peaceful interethnic co-existence.I have touched on only one of several conditions for ethnic survival and revival: the shared historical myths. Research in this field is essential if we are to begin to understand, and, thus, perhaps to alleviate, the many social and political problems in this area.Greet Van de Vyver is Aspirant of the National Foundation for Scientific Research of Belgium and a Ph.D. Candidate in the Department of Social and Cultural Anthropology, Catholic University of Leuven in Belgium.     

Mechanism of inhibition of mitochondrial enzymatic complex I-III by adriamycin derivatives

We demonstrate here that complex I-III of bovine heart mitochondrial membrane is inhibited by adriamycin derivatives. This inhibition is a cardiolipin-dependent process. This lipid, specific to the inner mitochondrial membrane, has been shown previously to interact specifically with adriamycin in model membranes (Goormaghtigh, E., Chatelain, P., Caspers, J. and Ruysschaert, J.-M. (1980) Biochim. Biophys. Acta 597, 1-14) and in mitochondrial membranes (Cheneval, D., Müller, M., Toni, R., Ruetz, S. and Carafoli, E. (1985) J. Biol. Chem. 260, 13003-13007). The differential scanning calorimetry data indicate that, in multilamellar liposomes, the formation of antibiotic-cardiolipin complexes induces a clustering of cardiolipin molecules. Conformational analysis of the antibiotic-cardiolipin complexes suggests that plane-plane interactions between the antibiotics aromatic moieties stabilize this complex formation. Possible mechanisms of inactivation of complex I-III by adriamycin are proposed.     

A CNBR peptide located in the middle region of diphtheria toxin fragment B induces conductance change in lipid bilayers: Possible role of an amphipathic helical segment

Conductance measurements on planar lipid bilayers demonstrate that CB1, a CNBr peptide of diphtheria toxin fragment B located in its middle region, possesses the unique property to destabilize the lipid bilayer organization. It is suggested that a segment of 25 amino acids in the N-terminal sequence of CB1 could be responsible for this effect. Its very low polarity, its predicted amphipathic helical structure and a helix length corresponding to the thickness of the hydrocarbon region of the lipid bilayer should specifically favor its insertion in the membrane. The existence of such a transverse lipid-associating domain could confer upon the molecule the properties leading to the anchoring of diphtheria toxin in the cytoplasmic membrane.     

Glucose induces membrane changes detected by fluorescence polarization in endocrine pancreatic cells

Pancreatic endocrine cells prepared from rat islets were labelled with 1,6-diphenyl-1,3,5-hexatriene and examined in a microviscosimeter. Glucose caused a dose-related decrease in fluorescence polarization. This decrease was observed within 1 min after increasing the concentration of glucose. These findings suggest that glucose affects membrane viscosity in pancreatic endocrine cells. At a glucose concentration of 16.7 mM, the estimated viscosity was 15 ± 3 per cent lower than basal value (2.01 ± 0.12 P).     

Functional differentiation of amphiphilic helices of the apolipoproteins by hydrophobic moment analysis

The amphiphilic character of different plasma apolipoproteins was investigated by a combination of established hydrophobicity analysis methods. These methods proved to be powerful in the detection of amphiphilic phospholipid-binding domains. Within this class of lipid-binding domains, lecithin-cholesterol acyltransferase activating and non-activating helices could be differentiated by calculating hydrophobic moments at different angles. We conclude that the hydrophobic characteristics of the different helices determined the mode of lipid binding and the substrate properties of these phospholipid-protein complexes for the lecithin-cholesterol acyltransferase reaction.     

Structural and functional asymmetry of the nucleotide-binding domains of P-glycoprotein investigated by attenuated total reflection Fourier transform infrared spectroscopy.

The dynamic changes occurring during the catalytic cycle of MDR3 P-glycoprotein (Pgp) and the role of each nucleotide-binding domain (NBD) in the transport process were investigated using attenuated total reflection Fourier transform infrared spectroscopy. For this purpose, wild-type Pgp and two mutations of homologous residues in each NBD were studied. On the one hand, we demonstrate here that, during its catalytic cycle, Pgp does not undergo secondary structure changes, but only modifications in its stability and accessibility to the external environment. On the other hand, amide H/D exchange kinetics demonstrate that homologous mutations in the two NBDs affect, in a different way, the dynamic properties of Pgp and also the dynamic changes occurring during ATP hydrolysis. These observations led to the conclusion that the NBDs have an asymmetric structure and different functions in the catalytic cycle of Pgp. Our data suggest that the release of drug from the membrane into the extracellular environment is due to decreased stability and/or increased accessibility to the external medium of the membrane-embedded drug-binding site(s). NBD1 would play an important role in this first restructuring of the membrane-embedded domains. NBD2 would be directly implicated in the subsequent restructuring of the membrane-embedded binding sites by which they recover their initial stability and accessibility to the membrane. It is proposed that this restructuring step would allow the binding and transport of another molecule of substrate.