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L T Boni

Publications and source records attributed to L T Boni.

26 records · Page 2Linked to original sources

Amphipathic helix and its relationship to the interaction of calcitonin with phospholipids.

Salmon, porcine, and human calcitonins interact with phosphatidylglycerol to form water-soluble complexes, but these peptides do not interact with the zwitterionic lipids phosphatidylcholine or sphingomyelin. The calcitonins are more helical in the presence of dimyristoylphosphatidylglycerol than in its absence, but human calcitonin is considerably less helical than the other two, particularly in the presence of the lipid. This may explain the previously reported faster rate of degradation of human compared with salmon calcitonin in vivo. The ability of human calcitonin to solubilize dimyristoylphosphatidylglycerol and to alter the phase transition properties of this phospholipid while maintaining a low content of helix indicates that the presence of an amphipathic helix is not a requirement for these effects. The binding of salmon calcitonin to dimyristoylphosphatidylglycerol has been studied by determining the dependence of the circular dichroism properties of the peptide on the concentration of lipid. At 25 degrees C, salmon calcitonin binds to five molecules of dimyristoylphosphatidylglycerol with an affinity constant of 1 X 10(5) M-1. Little change in these parameters is observed at 38 degrees C, and the complex is stable over a wide range of temperatures both above and below the phase transition temperature. The rate of reaction of salmon calcitonin with dimyristoylphosphatidylglycerol is rapid at or above the phase transition temperature of the lipid but not at low temperatures. Salmon calcitonin also interacts with egg phosphatidylglycerol. These results demonstrate that salmon calcitonin can react with phosphatidylglycerol at or above its phase transition temperature to form complexes which are at least kinetically stable both above and below the phase transition temperature. Salmon calcitonin can solubilize mixtures of dimyristoylphosphatidylglycerol and dimyristoylphosphatidylcholine containing 25% or more of the former phospholipid. The helical content of the peptide in the presence of these lipid mixtures is dependent on the fraction of the lipid which is phosphatidylglycerol, with larger fractions of this lipid leading to the formation of a higher helical content. At 25% phosphatidylglycerol, salmon calcitonin can solubilize the lipid mixture without much increase in the helix content of the peptide, again demonstrating that an amphipathic helical structure is not required for the solubilization of phospholipids. Ionic bonding appears to be an important component in the binding of the cationic calcitonins to phospholipids. Salmon calcitonin binds to the acidic phospholipids phosphatidylinositol and phosphatidic acid, but not to zwitterionic phospholipids. In addition, high concentrations of NaCl cause the dissociation of the complex between salmon calcitonin and dimyristoylphosphatidylglycerol.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Polymorphic phase behaviour of dilinoleoylphosphatidylethanolamine and palmitoyloleoylphosphatidylcholine mixtures. Structural changes between hexagonal, cubic and bilayer phases.

The polymorphic phase behaviour of dilinoleoylphosphatidyethanolamine (DLPE) and 1-palmitoyl-2-oleoylphosphatidylcholine (POPC) is investigated by freeze-fracture electron microscopy, X-ray diffraction and 31P-NMR. The structures at 5% or less POPC are predominantly inverted hexagonal (HII), whereas at 15% or more POPC, the structure is mostly bilayer (L), interrupted by defects (lipidic particles). A cubic phase structure is observed in the transition range between H and L phases; the cubic arrangement deteriorates at higher temperatures into an amorphous aggregate of spherical units. Both cubic and amorphous structures contribute to the isotropic 31P resonance, with no preference for PC or PE partitioning in the isotropic motion as observed by high resolution NMR. The existence of the cubic phase seems to depend cirtically on the homogeneity and the degree unsaturation of the phospholipids.

Freeze Fracturing↗

The nature of lipidic particles and their roles in polymorphic transitions.

The structural transition between bilayer (L alpha), inverted hexagonal (HII) and inverted cubic (CII) phases in mixtures of unsaturated phosphatidylethanolamines (PE) and phosphatidylcholines (PC) were investigated. Freeze fracture electron micrographs of intermediate stages of phase transitions showed that CII was a stable intermediate form between the L alpha--HII transition. The electron microscopic observation was supported by X-ray diffraction and 31P-NMR results. Detailed morphology revealed that during the L alpha--CII transition, interlamellae attachment points (conical lipidic particles) connect adjacent bilayers to form arrays of entrapped water pockets (inverted micelles). These water-containing spherical units were packed in a cubic lattice. In the CII to HII transition, these spherical units were linearly connected to form tubes. During the L alpha--HII transition, a ripple pattern was observed across the otherwise smooth lamellar. The troughs of the ripples were transformed into linear connections between adjacent bilayers, thereby converting multilayer structures into parallel tubes. No lipidic particles were involved in this type of transition. We show that there are different mechanisms involved in the L alpha, HII, CII polymorphic transitions, and that different types of 'lipidic particles' representing different molecular organizations may be involved in each case. Models of these transitions are proposed.

Lipid Bilayers↗

Membrane fusion through point defects in bilayers.

Fusion between bilayers of mixed egg phosphatidylcholine and soybean phosphatidylethanolamine was induced by freezing and thawing. Contact points between bilayers were observed by freeze fracture electron microscopy, and isotropic molecular motional averaging was detected by phosphorus-31 nuclear magnetic resonance under fusion conditions. A molecular model of point defect structure is proposed as an intermediate stage of fusion.

Freeze Fracturing↗

Lipid-polyethylene glycol interactions: I. Induction of fusion between liposomes.

Fusion between unilamellar vesicles of both egg phosphatidylcholine and bovine phosphatidylserine was induced by polyethylene glycol. Aggregation and fusion events were monitored by electron microscopy and turbidity measurements. The threshold concentration of polyethylene glycol for aggregation and fusion is found to be independent of lipid concentration. Typically, aggregation of phosphatidylcholine vesicles starts at 2.5% (wt/wt) polyethylene glycol, but fusion is not significant until the polyethylene glycol concentration reaches 35%. Multilamellar vesicles were formed as a result of fusion.

Animals↗

Lipid-polyethylene glycol interactions: II. Formation of defects in bilayers.

The kinetic properties of L-leucine transport across the human red blood cell membrane was analyzed according to the simple pore and carrier theory of Lieb and Stein (Biochim. Biophys. Acta, 1974, 373: 165-177 and 178-196) at 25 degrees C, pH 7.4. Several methods were used in order to obtain a thorough kinetic description of L-leucine transport. A rejection of the simple pore model was suggested from the result of zero-trans influx and zero-trans and equilibrium-exchange efflux experiments. Several predictions from the simple carrier model, based on the requirement of consistency among different kinetic parameters, were tested in infinite experiments, i.e. experiments performed at a high concentration of substrate at one of the faces of the membrane. The simple pore model was rejected, but no crucial evidence against a simple carrier model, which displays symmetric properties at 25 degrees C, was found in the concentration range considered (0.002-68 mM). The relative magnitudes of the rate constants of the translocation process are discussed, and it is concluded (a) that both the dissociation and translocation of carrier-complex is faster than the translocation of the empty carrier, (b) that no translocation step is rate determining, and (c) that the carrier-complex is equally distributed across the membrane at equilibrium. The present work provides a unique example of a carrier-mediated transport mechanism which displays symmetric properties. L-leucine transport in red blood cells may be a convenient system for studying molecular mechanisms of facilitated transport.

Cell Fusion↗