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V Luzzati

Publications and source records attributed to V Luzzati.

At least 19 recordsLinked to original sources
PubMed1995

Topological properties of two cubic phases of a phospholipid:cholesterol:diacylglycerol aqueous system and their possible implications in the phospholipase C-induced liposome fusion.

Water dispersions of phospholipid:cholesterol:diacylglycerol may, under certain conditions, originate either the lipid- and water-permeable Q224 cubic phase, or the lipid-permeable but water-impermeable Q227 cubic phase. These results are discussed within the framework of the phospholipase C-induced fusion of liposomes [Nieva et al. (1993) Biochemistry 32, 1054]. It is suggested that the cubic phases Q224 and Q227 represent two classes of lipid organization, one promoting, the other hindering the mixing of aqueous contents that is characteristic of membrane fusion. In this context, inverted micelles appear to be the end point of the fusion process, rather than fusion intermediates.

Bacillus cereus↗
PubMed1995

Order-disorder phenomena in myelinated nerve sheaths: V. Effects of temperature on rat sciatic and optic nerves, and structural differences between the two types of nerve.

We describe in this work X-ray scattering and electron microscope studies of rat sciatic and optic nerves as a function of temperature. The scattering experiments were analyzed as described in the previous papers of this series: a variety of parameters were determined, some of which characterize the lattice disorder, others the structure of the motif. The main results are the following. All the parameters determined by the X-ray scattering study vary with temperature and the temperature-dependence is specific for the type of nerve (sciatic or optic). Most of the disorder-related parameters display a minimum or a maximum in the vicinity of physiological temperature (38 degrees C in rat); this observation, strongly supported by the electron microscope study, shows that the degree of organization of myelin is highest near physiological temperature. The structure of the motif, as revealed by the electron density profile, is fairly different in the two types of nerves (in contrast with the assumption made by previous workers); the structure also varies with temperature and the temperature-induced alterations are nerve-type specific. In the two types of nerve the thickness of the lipid bilayer varies with temperature as expected for a lipid-containing system with hydrocarbon chains in the disordered conformation. In sciatic nerve the thickness of the (thinner) cytoplasmic polar layer, which is also the layer most affected by lattice disorder in this type of nerve, decreases dramatically with increasing temperature. In optic nerve, in which lattice disorder predominantly affects the extracellular layer, the thickness of both the cytoplasmic and the extracellular layer is barely affected by temperature.

Algorithms↗
PubMed1992

Order-disorder phenomena in myelinated nerve sheaths. IV. The disordering effects of high levels of local anaesthetics on rat sciatic and optic nerves.

Sequences of 15 minute X-ray scattering spectra were recorded with rat sciatic and optic nerves, superfused with tetracaine-containing Ringer solutions. The spectra were analysed using the algorithm advocated in this series of papers. The main results, as a function of the time of exposure to tetracaine, were: the mean value of the repeat distance increases; its variance decreases; the average number of membrane pairs per coherent domain decreases; the fraction of isolated membrane pairs increases. Eventually, the spectra were observed to give way to the continuous intensity curve of a single, isolated membrane pair. At all stages of the experiment the continuous intensity curves were found to differ from one type of nerve to the other, and to be invariant, for each type of nerve, with respect to the tetracaine treatment. The X-ray scattering study clearly identified the nature of the structural differences between the two types of myelin sheaths: in that of native sciatic nerves, packing disorder preferentially affects the cytoplasmic space of the membrane pair, and tetracaine disrupts the packing in that space; in the myelin of optic nerves it is the external space that is preferentially affected by packing disorder and disrupted by tetracaine. The time-course of the structure parameters showed that, at any stage of the experiment, tetracaine acts preferentially on the more highly disordered regions of the structure and totally disrupts them. These results corroborate earlier conclusions reported in the previous papers of this series. An electron microscope study was also performed on tetracaine-treated nerves: the results, in close agreement with those of the X-ray scattering study, neatly confirm the conclusions given above. In a more general way, the remarkable agreement between the results of the analysis of the X-ray scattering spectra and the electron microscope observations strongly supports the validity of the physical model used in this series of papers and the correctness of the mathematical treatment that we advocate. Finally, the relations between this work and the work of others are discussed. It must be stressed that the present work bears on the toxic rather than on the anaesthetic effects of tetracaine.

Action Potentials↗
PubMed1992

Cubic phases of lipid-containing systems. The structure of phase Q223 (space group Pm3n). An X-ray scattering study.

The hexagonal (H) and the cubic (Q223) phases of the systems dodecyltrimethylammonium chloride-water and palmitoyllysophosphatidy choline-water have been studied by X-ray scattering techniques. The signs of the reflections of phase H were determined by a systematic study as a function of the water content, those of phase Q223 were assessed using a pattern recognition approach based upon the axiom that the histograms of the electron density maps of phases Q223 and H, extrapolated to the same concentration and properly normalized in scale and shape, are very similar to each other. In the case of phase Q223, all the sign combinations (the phi-sets) compatible with the observed reflections were generated, and each of the corresponding histograms was compared with the histogram of the map of phase H. One novelty of this work is the use of a highly sensitive criterion to estimate the similarity of the histograms, namely the distance in the six-dimensional space of the moments [mean value of (delta rho)n]1/n, for 3 greater than or equal to n greater than or equal to 8. In the two systems, the use of this criterion has led to the unambiguous choice of one electron density map. The maps show that the structure of phase Q223 consists of disjointed micelles (of type I), belonging to two different classes: those of one class are quasi-spherical in shape and are centered at the points a, those of the other class are disc-shaped and are centred at the points c. The results of this work rule out a structure formed by a cage-like distribution of rods enclosing a set of quasi-spherical micelles and is consistent with previous proposals. This is the second example, after that of phase Q227, of a micellar cubic phases in lipid-containing systems; all the known examples of phase Q223 are of type I, those of phase Q227 of type II.

Freeze Fracturing↗
PubMed1992

Lipid polymorphism: a correction. The structure of the cubic phase of extinction symbol Fd-- consists of two types of disjointed reverse micelles embedded in a three-dimensional hydrocarbon matrix.

The X-ray scattering study of a cubic phase of extinction symbol Fd--, recently performed on a lipid extract (PFL) from Pseudomonas fluorescens [Mariani et al. (1990) Biochemistry 29, 6799-6810] has been extended to several other systems, all consisting of mixtures of water-miscible (MO, PC, PE, oleate) and of water-immiscible (FA, DG) lipids, plus water. In all of these systems the cubic phase was observed in the presence of excess water. Some inconsistencies observed between PFL and the other systems, the fact that in PFL one of the reflections of the cubic phase happened to coincide with the strongest reflection of the hexagonal phase, and the finding, in one of the original cubic samples of PFL kept in the cold for more than 3 years, that the intensity of one of the reflections had decreased dramatically all indicated that a nonnegligible amount of a hexagonal impurity was in fact present in the samples of PFL originally thought to contain a pure cubic phase. The intensities were corrected for that impurity and analyzed again using a pattern recognition approach based upon the axiom that the histogram of the electron density maps is invariant with respect to physical structure, when different phases are compared whose chemical composition is the same. The hexagonal phase provided the reference phase for the comparison. The moments mean value of (delta rho)n were used to compare the histograms.(ABSTRACT TRUNCATED AT 250 WORDS)

Crystallography↗
PubMed1991

Order-disorder phenomena in myelinated nerve sheaths. III. The structure of myelin in rat optic nerves over the course of myelinogenesis.

An X-ray scattering study was performed on optic nerves dissected from rats aged from ten days to one year. The spectra were analysed using the procedure described in the previous papers of this series. Each experiment yields the values of a variety of parameters: the average D and the variance sigma D of the repeat distance, the average number mean value of N of motifs per crystallite, the fraction alpha loose of myelin that does not belong to the compact sheaths, the sets [idiff(h/D)] and [imotif(k/2D)] that suffice to define, respectively, the spurious scattering and the continuous intensity curve of the elementary membrane pair. A surprising result is that, in the native optic, as previously found in the swollen sciatic nerves, the stacking disorder affects the external space, whereas in native sciatic nerves the disorder affects the cytoplasmic space. An analysis of the evolution of the structure parameters as a function of the age of the animal and a comparison with the results previously obtained with rat sciatic nerves led to the following conclusions: the structure of the elementary membrane pair is constant throughout myelinogenesis; mean value of N is much smaller in optic than in sciatic nerves; mean value of N and the degree of myelination increase with age in the two types of nerve; D is smaller in optic than in sciatic nerves; in optic nerves, D decreases slightly with age, but in sciatic nerves it increases; sigma D is strongly age-dependent in optic nerves, but almost age-independent in sciatic nerves. In contrast to sciatic, the structure of optic nerve myelin was found to be almost insensitive to hypertonic solutions. Finally, a pair of electron density profiles was selected, quite similar to those selected previously in sciatic nerves, one corresponding to Caspar & Kirschner's the other to Worthington & McIntosh's proposals, neither of which can be ruled out according to the criteria used in this work.

Aging↗
PubMed1990

Order-disorder phenomena in myelinated nerve sheaths. I. A physical model and its parametrization: exact and approximate determination of the parameters.

An algorithm is developed for the analysis of the X-ray scattering spectra of lamellar systems, by reference to a precise physical model. The model consists of identical planar lamellae (the motif), all parallel and stacked in a one-dimensional crystal with four types of defect: stacking disorder, finite size of the crystallites, and presence of diffuse and blank scattering. In addition, the spectra are distorted by collimation aberrations. In order to evaluate the effects of these distortions, the following assumptions are made: (1) beyond some point Slimit the intensity curve can be expressed as a function of a (small) number of parameters; (2) the blank scattering, restricted to very small angles, can be identified and eliminated; and (3) the diffuse scattering is entirely defined by the values of idiff(h/D) at the lattice Sh = h/D (h is a positive integer less than or equal to DSlimit). These assumptions lead to an expression of the whole of the intensity curve as a function of a finite number of parameters: the average D and the variance sigma 2D of the repeat distance, the average number [N] of lamellae per crystallite, the set [idiff(h/D)] and the set [imotif(k/2D)] (where k is a positive integer), which defines the structure of the motif. An algorithm is proposed to determine the value of the various parameters. The derivation of the algorithm involves several operations: construction in real space of periodic functions whose motifs are step-sections of the autocorrelation function; expression in reciprocal space, and in terms of the experimental scattering curves, of the Fourier transform of those periodic functions; analysis of the properties of the two functions. The algorithm is tested using a variety of simulated scattering curves whose parameters [imotif(k/2D)], [idiff(2/D)], D, sigma D, [N] (and collimation distortions) are within the range commonly encountered in experimental conditions. The results show that the values of the parameters retrieved by the algorithms are very close to those used in the simulation. The calculations are fast and easy to implement on a computer. The main virtues of the algorithm are (1) to determine the values of all the parameters at once, eliminating most of the intermediate (and questionable) manipulations (separation of signal from noise, discrimination of overlapping reflections, integration of the intensities) and (2) to yield the continuous intensity curve of a single motif.

Algorithms↗
PubMed1990

Order-disorder phenomena in myelinated nerve sheaths. II. The structure of myelin in native and swollen rat sciatic nerves and in the course of myelinogenesis.

The algorithm described in the accompanying paper was applied to X-ray scattering experiments performed with rat sciatic nerves, either as a function of the age of the animal (4 to 30 days), or with adult nerves swollen in non-isotonic media. The results were all consistent with the model of disorder used in the theoretical treatment. The algorithm leads, in one step, from the data to the numerical values of the parameters, avoiding all intermediate manipulation. For each experiment a variety of parameters was determined: the average D and the variance sigma 2D of the repeat distance, the average number [N] of motifs per crystallite, the set [idiff(h/D)], which defines the diffuse scattering, the fraction alphaloose of myelin that does not belong to the compact sheaths, and the set [imotif (k/2D)], which suffices to define the continuous intensity curve of the motif imotif(s). Note the remarkable wealth of information, especially by contrast with conventional analyses which, as a rule, only yield the values of D and of the set [imotif(h/D)] (insufficient to determine the function imotif(s]. The function imotif(s) and the parameters D and sigma D (and thus the local structure of the myelin sheaths) were shown to be almost invariant in the course of myelinogenesis; what varies is mainly the total amount of myelin in the nerve and the number of membranes per sheath. Swelling agents have a dramatic influence on the X-ray scattering spectra, but in spite of the conspicuous variation of D, sigma D and [N] the structure of the motif is invariant. The structure of the motif was shown to be quite different in the native and in the swollen samples; the stacking disorder appears to involve mainly the cytoplasmic space in native myelin, the external space in swollen nerves. The very notion of electron density profile, when disorder is present, is discussed. Two criteria were proposed to select the "best" signs of the reflections: two sets came out at almost the same rank, one corresponding to Caspar & Kirschner's the other to Worthington & McIntosh's proposals, neither of which can be ruled out according to the criteria used in this work.

Algorithms↗
PubMed1990

Polymorphism of a lipid extract from Pseudomonas fluorescens: structure analysis of a hexagonal phase and of a novel cubic phase of extinction symbol Fd--.

The phase diagram of the Pseudomonas fluorescens lipid extract is unusual, in the sense that it displays a cubic phase straddled by a hexagonal phase. The hexagonal phase was studied over an extended concentration range, and the reflections were phased on the assumption that the structure contains circular cylinders of known radius. The cubic phase, whose extinction symbol is Fd--, was analyzed by reference to space group No. 227 (Fd3m). The phases of the reflections were determined by using a novel pattern recognition approach, based upon the notion that the average fourth power of the electron density contrast mean value of (delta r)4 is dependent on chemical composition but not on physical structure, provided that the function delta r(r) satisfies the constraints mean value of (delta r) = 0 and mean value of (delta r)2 = 1. As a further constraint, a shape normalization is used, in the form of a Gaussian apodization of the intensities, which has the effect of normalizing the curvature of the autocorrelation function at the origin. We analyzed two cubic samples of different composition: for each of them we generated all the phase combinations compatible with the X-ray scattering data and we searched for those whose mean value of (delta r)4 best agrees with the hexagonal phase. Taking advantage of the favorable properties of the phase diagram, we carefully explored the effects of various parameters; we concluded that the chemical composition of the phases being compared must be identical, that the X-ray scattering data should not be truncated artificially, and that the apodization must be mild so that the curvature takes a value intermediate between those corresponding to the raw data of the two phases. When all these precautions were taken, mean value of (delta r)4 was found to be remarkably invariant; this conclusion is important in view of the possible usefulness of the novel technique in tackling ab initio--and at very low resolution--structural problems of more general interest. The structure of the cubic phase consists of a 3D network of rods joined tetrahedrally 4 by 4 according to a diamond lattice and of a family of quasi-spherical disjointed micelles; the core of the rods and of the micelles is polar, and the interstices are filled by the hydrocarbon chains (structure of type II). All the dimensions (diameter of rods and micelles, area per chain at the polar/apolar interface) are consistent with the chemical properties of the system.(ABSTRACT TRUNCATED AT 400 WORDS)

Lipids↗
PubMed1988

Cubic phases of lipid-containing systems. Structure analysis and biological implications.

Of the six cubic phases identified so far in lipid-containing systems, the structures of only two have been determined unambiguously. We tackle the structure determination of the other four. We use for that purpose a novel pattern recognition approach, which consists of generating all the sets of phase angles (phi-sets) compatible with the observed reflections, and of screening them in a search for the "best" one. Two criteria are used for screening: both involve the parameter [(delta rho)4] (delta rho is a dimensionless function proportional to the Fourier transform of the set of observed structure factors). One is a test of smoothness, based upon the postulate that the "best" phi-set is that whose [(delta rho)4] is minimum; this criterion, equivalent to maximum entropy, is fulfilled when the system is devoid of heavy atoms, and when the polar and the hydrocarbon moieties occupy almost equal volumes. The other criterion is based upon the notion that [(delta rho)4] takes the same (or similar) values in thermodynamic phases with the same (or similar) chemical composition, whatever the structure of the phases. The validity of the two criteria is verified using numerous examples. The six cubic phases are analysed using this approach. The structure of three of them (Q230, Q224, Q229) can be described in terms of two three-dimensional networks of connected rods, mutually intertwined and unconnected: in Q230 the rods are coplanarly joined 3 by 3; in Q224 the rods are tetrahedrally joined 4 by 4; in Q229 the rods are cubically joined 6 by 6. The structures of Q212 and Q227 are related to those of Q230 and Q224, respectively; one of the two networks of rods is preserved, the other is replaced by a lattice of closed micelles. The structure of Q223 appears to consist of a cage-like continuous three-dimensional network of connected globules, coplanarly joined 3 by 3 at one end and 4 by 4 at the other, enclosing a three-dimensional lattice of closed micelles. The analogies of the structures of Q230, Q224, Q229 with the three fundamental cubic infinite periodic minimal surfaces are discussed. More interestingly, the structures of, on the one hand Q230, Q224, Q229 and of Q212, Q227, Q223 on the other, are shown to provide topological generalizations of the two paradigms of lipid organization; namely, the bilayer and the monolayer.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1988

Tetraether lipid components from a thermoacidophilic archaebacterium. Chemical structure and physical polymorphism.

As a continuation of an X-ray scattering study of the tetraether lipids extracted from the thermophilic archaebacterium Sulfolobus solfataricus, the phase behaviour of four fractions of the complex polar lipid extract (PLE) is described. Each molecule of two of these fractions (P1 and GL) carries an unsubstituted glycerol headgroup, those of another (P2) no such group; the fourth fraction (WPLE) is obtained by water-washing PLE, thus reducing its P2 content from approximately 48% to approximately 24% and increasing the average number of molecules bearing an unsubstituted glycerol headgroup from approximately 0.4 to approximately 0.6. The main result is a striking correlation between the phase behaviour and the average ratio of unsubstituted glycerol headgroups to the total number of headgroups: the fractions P1, GL and WPLE, in which that number is respectively 0.5, 0.5 and 0.3, form rod-containing phases; the fraction P2, in which that number is zero, yields a lamellar phase throughout the phase diagram. An analysis of the dimensions of the structure elements confirms our previous conclusion that, in the presence of a sufficient amount of water, the unsubstituted glycerol headgroups partition preferentially in the hydrocarbon regions rather than at the polar/apolar interfaces. These results, moreover, corroborate our previous conjectures regarding the correlations between the structure of the plasma membrane, the phase behaviour of the lipid extract and life at high temperature.

Archaea↗
PubMed1985

Structure and polymorphism of bipolar isopranyl ether lipids from archaebacteria.

We describe in this work the structure and polymorphism of a variety of lipids extracted from Sulfolobus solfataricus, an extreme thermoacidophilic archaebacterium growing at about 85 degrees C and pH 2. These lipids are quite different from the usual fatty acid lipids of eukaryotes and prokaryotes: each molecule consists of two C40 omega-omega' biphytanyl residues (with 0 to 4 cyclopentane groups per residue), ether linked at both ends to two (variably substituted) glycerol or nonitol groups. Four lipid preparations were studied; the total and the polar lipid extracts, and two hydrolytic fractions, the symmetric glycerol dialkyl glycerol tetraether and the asymmetric glycerol dialkyl nonitol tetraether, as a function of water content and temperature, using X-ray scattering techniques. The main conclusions from the study of the four lipid preparations can be summarized as follows. (1) As with other lipids, a remarkable number and variety of phases are observed over a temperature-concentration range close to "physiological" conditions. The possibility is discussed that this polymorphism reflects a fundamental property of lipids, closely related to their physiological rôle. (2) As in other lipids, two types of chain conformations are observed: a disordered one (type alpha) at high temperature; at lower temperature, a more ordered packing of stiff chains, all parallel to each other (type beta'). At temperatures and degrees of hydration approaching the conditions prevailing in the living cell, the conformation is of type alpha. (3) In all the phases with chains in the alpha conformation, the unsubstituted glycerol headgroups, whose concentration is high in these lipids, segregate in the hydrocarbon matrix, away from the other polar groups. This property may have interesting biological consequences: for example, the chains of a fraction of the bipolar lipid molecules can span hydrocarbon gaps as wide as 75 A. (4) Two cubic phases are observed in the total and the polar lipid extracts, which display a remarkable degree of metastability, most unusual in lipid phase transitions involving structures with chains in the alpha conformation. This phenomenon can be explained by the interplay of the physical structure of the cubic phases (the two contain two intertwined and unconnected three-dimensional networks of rods) and the chemical structure of the lipid molecules: the two headgroups of most molecules being anchored on each of the two networks of rods, the migration of the lipid molecules is hindered by the two independent diffusion processes and by the entanglement of the chains.(ABSTRACT TRUNCATED AT 400 WORDS)

Archaea↗
PubMed1981

Thinking about the possible role of lipids.

We contemplate the possibility that lipids play a subtler functional role in membranes than that of providing a permeability barrier and a 2-dimensional solvent for proteins. An analysis of the structural properties of lipids, as established by X-ray scattering studies of model systems, suggests a variety of molecular processes involving the structure of the lipid matrix in the functions of membranes. By way of illustration, a hypothetical structural transition is considered, involving nerve membrane lipids and underlying the action potential. The design of an experiment, aimed at testing this hypothesis, is described.

Action Potentials↗
PubMed1978

The influence of protein-lipid interactions on the order-disorder conformational transitions of the hydrocarbon chain.

The phases of simple systems involving one type of protein (lysozyme or cytochrome c) and one type of lipid (phosphatidic acid) have been characterized by X-ray crystallography, chemical analysis and spin-labeling technique as a function of temperature. They are of the lamellar type with alternative protein monolayers and lipid bilayers. According to the pH, two types of lamellar phases are obtained, one where the lipid-protein interactions are mainly hydrophobic, the other where they are electrostatic. In both cases, a phase transition occurs as temperature is lowered, between a high temperature phase, where all the lipids are in the liquid-like state, and another phase where some lipid chains are rigid. In the case of the phases with electrostatic interaction, it is shown that the onset of the order-disorder transition is shifted towards low temperature as compared with the homologous lipid-water phase and that the protein content of the phase decreases as the ratio of the liquid to rigid hydrocarbon chains decreases. This leads us to suggest that in the systems studied in this work the proteins interact only with lipid in the liquid-like state. In the case of the phases with hydrophobic interaction, it is shown that the extent of hydrophobic interaction between protein and lipid increases as the unsaturation of the hydrocarbon chains increases. The onset of the order-disorder transition shows a greater shift towards low temperature than the one observed in the case of the phase with electrostatic interaction.

Cytochrome c Group↗
PubMed1977

A novel packing of the hydrocarbon chains in lipids. The low temperature phases of dipalmitoyl phosphatidyl-glycerol.

The system dipalmitoyl phosphatidylglycerol-water displays several phases in the temperature-concentration range explored in this work. All the phases are lamellar; they differ by the organization of the hydrocarbon chains. In the high temperature phase the conformation of the chains is liquid-like. In the low temperature phases the chains are stiff and parallel and they interdigitate (in other words the CH3 ends of the chains of one layer are near to the polar groups of the opposite layer). Moreover, several types of packings of the stiff chains are observed which differ by the symmetry of the two-dimensional lattices. The observed lattices are p6, cmm and pgg.

Molecular Conformation↗