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J Katsaras

Publications and source records attributed to J Katsaras.

At least 19 recordsLinked to original sources

Preferential accumulation of Abeta(1-42) on gel phase domains of lipid bilayers: an AFM and fluorescence study.

Peptide-membrane interactions have been implicated in both the toxicity and aggregation of beta-amyloid (Abeta) peptides. Recent studies have provided evidence for the involvement of liquid-ordered membrane domains known as lipid rafts in the formation and aggregation of Abeta. As a model, we have examined the interaction of Abeta(1-42) with phase separated DOPC/DPPC lipid bilayers using a combination of atomic force microscopy (AFM) and total internal reflection fluorescence microscopy (TIRF). AFM images show that addition of Abeta to preformed supported bilayers leads to accumulation of small peptide aggregates exclusively on the gel phase DPPC domains. Initial aggregates are observed approximately 90 min after peptide addition and increase in diameter to 45-150 nm within 24 h. TIRF studies with a mixture of Abeta and Abeta-Fl demonstrate that accumulation of the peptide on the gel phase domains occurs as early as 15 min after Abeta addition and is maintained for over 24 h. By contrast, Abeta is randomly distributed throughout both fluid and gel phases when the peptide is reconstituted into DOPC/DPPC vesicles prior to formation of a supported bilayer. The preferential accumulation of Abeta on DPPC domains suggests that rigid domains may act as platforms to concentrate peptide and enhance its aggregation and may be relevant to the postulated involvement of lipid rafts in modulating Abeta activity in vivo.

1,2-Dipalmitoylphosphatidylcholine↗

Detection of submicron-sized raft-like domains in membranes by small-angle neutron scattering.

Using coarse grained models of heterogeneous vesicles we demonstrate the potential for small-angle neutron scattering (SANS) to detect and distinguish between two different categories of lateral segregation: 1) unilamellar vesicles (ULV) containing a single domain and 2) the formation of several small domains or "clusters" (approximately 10 nm in radius) on a ULV. Exploiting the unique sensitivity of neutron scattering to differences between hydrogen and deuterium, we show that the liquid ordered (lo) DPPC-rich phase can be selectively labeled using chain deuterated dipalymitoyl phosphatidylcholine (dDPPC), which greatly facilitates the use of SANS to detect membrane domains. SANS experiments are then performed in order to detect and characterize, on nanometer length scales, lateral heterogeneities, or so-called "rafts", in approximately 30 nm radius low polydispersity ULV made up of ternary mixtures of phospholipids and cholesterol. For 1:1:1 DOPC:DPPC:cholesterol (DDC) ULV we find evidence for the formation of lateral heterogeneities on cooling below 30 degrees C. These heterogeneities do not appear when DOPC is replaced by SOPC. Fits to the experimental data using coarse grained models show that, at room temperature, DDC ULV each exhibit approximately 30 domains with average radii of approximately 10 nm.

1,2-Dipalmitoylphosphatidylcholine↗

Finite-size effects in biomimetic smectic films.

Thin stacks of lipid multibilayers supported on rigid silicon and mica substrates are found to exhibit finite-size effects. Using neutron diffraction we find that the repeat spacing (d) of stacks containing up to a few tens of bilayers depends on their thickness (D), with d increasing with decreasing D. Differences in d are larger in the low-temperature Lbeta' phase consisting of rigid bilayers than in the high-temperature Lalpha phase where the bilayers are more flexible. Various scenarios that may be responsible for this counterintuitive observation are discussed.

Biomimetic Materials↗

Relationship between the unbinding and main transition temperatures of phospholipid bilayers under pressure.

Using neutron diffraction and a specially constructed high pressure cell suitable for aligned multibilayer systems, we have studied, as a function of pressure, the much observed anomalous swelling regime in dimyristoyl- and dilauroyl-phosphatidylcholine bilayers, DMPC and DLPC, respectively. We have also reanalyzed data from a number of previously published experiments and have arrived at the following conclusions. (a). The power law behavior describing anomalous swelling is preserved in all PC bilayers up to a hydrostatic pressure of 240 MPa. (b). As a function of increasing pressure there is a concomitant decrease in the anomalous swelling of DMPC bilayers. (c). For PC lipids with hydrocarbon chains >or=13 carbons the theoretical unbinding transition temperature T small star, filled is coupled to the main gel-to-liquid crystalline transition temperature T(M). (d). DLPC is intrinsically different from the other lipids studied in that its T small star, filled is not coupled to T(M). (e). For DLPC bilayers we predict a hydrostatic pressure (>290 MPa) where unbinding may occur.

Binding Sites↗

Spontaneously formed monodisperse biomimetic unilamellar vesicles: the effect of charge, dilution, and time.

Using small-angle neutron scattering and dynamic light scattering, we have constructed partial structural phase diagrams of lipid mixtures composed of the phosphatidylcholines dimyristoyl and dihexanoyl doped with calcium ions (Ca2+) and/or the negatively charged lipid, dimyristoyl phosphatidylglycerol (DMPG). For dilute solutions (lipid concentration < or =1 wt %), spontaneously forming unilamellar vesicles (ULVs) were found, and their polydispersity was determined to be approximately 20%. The stability of the Ca2+- or DMPG-doped ULVs was monitored over a period of 4 days and their structural parameters (e.g., average outer radius, ) were found to be insensitive to the lipid concentration (Clp). However, doping the dimyristoyl/dihexanoyl system with both Ca2+ and DMPG resulted in ULVs whose was found to be Clp dependent. The of DMPG-doped ULVs remained unchanged over an extended period of time (at least 4 days), a good indication of their stability.

Calcium↗

Finite-size effects do not reduce the repeat spacing of phospholipid multibilayer stacks on a rigid substrate.

Finite-size effects in stacks of phospholipid bilayers, in the fluid L alpha phase, are investigated using samples oriented on silicon substrates. Recently in this journal, such effects have been suggested as the probable cause of reduced lamellar repeat spacings in very thin samples made up of a few (<10) bilayers. Our systematic studies on samples of different thicknesses do not support this conclusion. At full hydration all samples are found to have the same repeat spacing, irrespective of their thickness. At lower hydrations, on the other hand, very thin samples, consisting of only a few bilayers, have a slightly larger spacing.

Biophysics↗

Concentration-independent spontaneously forming biomimetric vesicles.

In this Letter we present small-angle neutron scattering data from a biomimetic system composed of the phospholipids dimyristoyl and dihexanoyl phosphorylcholine (DMPC and DHPC, respectively). Doping DMPC-DHPC multilamellar vesicles with either the negatively charged lipid dimyristoyl phosphorylglycerol (DMPG, net charge -1) or the divalent cation, calcium (Ca2+), leads to the spontaneous formation of energetically stabilized monodisperse unilamellar vesicles whose radii are concentration independent and in contrast with previous experimental observations.

Biomimetic Materials↗

Discontinuous unbinding of lipid multibilayers.

We have observed a discontinuous unbinding transition of lipid bilayer stacks composed of phosphatidylethanolamine and phosphatidylglycerol using x-ray diffraction. The unbinding is reversible and coincides with the main (L(beta)-->L(alpha)) transition of the lipid mixture. Interbilayer interaction potentials deduced from the diffraction data reveal that the bilayers in the L(beta) phase are only weakly bound. The unbinding transition appears to be driven by an abrupt increase in steric repulsion resulting from increased thermal undulations of the bilayers upon entering the fluid L(alpha) phase.

Lipid Bilayers↗

Kinetic pathway of the bilayered-micelle to perforated-lamellae transition.

Using time-resolved small-angle neutron scattering, we have studied the kinetics of the recently observed bilayered-micelle (or so-called "bicelle") to perforated-lamellar transition in phospholipid mixtures. The data suggest that phase-ordering occurs via the early-time coalescence of bicelles into stacks of lamellae that then swell. Our measurements on this biomimetic system highlight the ubiquitous role of transient metastable states in the phase ordering of complex fluids.

Biophysics↗

Enhancement of steric repulsion with temperature in oriented lipid multilayers.

We have studied the temperature dependence of the stacking periodicity, d, of oriented phospholipid multilayers using grazing angle neutron scattering techniques. d is found to increase substantially at higher temperatures, just before the bilayers peel off from the substrate. Although we do not observe thermal unbinding, our results are consistent with the notion that the unbinding transition is driven by steric repulsion arising from thermal fluctuations of the membranes, in contrast to those of a recent study by Vogel et al. [Phys. Rev. Lett. 84, 390 (2000)].

Dimyristoylphosphatidylcholine↗

Observation of Kossel and Kikuchi lines in thermal neutron incoherent scattering.

In this Letter we report the observation of K lines (representing collectively, Kossel and Kikuchi lines) produced by monochromatic thermal neutrons interacting with a KDP (potassium dihydrogen phosphate) single crystal. Since K lines contain phase information, these observations establish the experimental basis for direct crystallographic phasing of atomic structures containing incoherent scatterers, such as hydrogen, via thermal neutron "inside source" holography.

Journal Article↗

Atomic structure holography using thermal neutrons.

The idea of atomic-resolution holography has its roots in the X-ray work of Bragg and in Gabor's electron interference microscope. Gabor's lensless microscope was not realized in his time, but over the past twelve years there has been a steady increase in the number of reports on atomic-resolution holography. All of this work involves the use of electrons or hard X-rays to produce the hologram. Neutrons are often unique among scattering probes in their interaction with materials: for example, the relative visibility of hydrogen and its isotopes is a great advantage in the study of polymers and biologically relevant materials. Recent work proposed that atomic-resolution holography could be achieved with thermal neutrons. Here we use monochromatic thermal neutrons, adopting the inside-source concept of Szöke, to image planes of oxygen atoms located above and below a single hydrogen atom in the oxide mineral simpsonite.

Journal Article↗

Morphology of fast-tumbling bicelles: a small angle neutron scattering and NMR study.

Bilayered micelles, or bicelles, which consist of a mixture of long- and short-chain phospholipids, are a popular model membrane system. Depending on composition, concentration, and temperature, bicelle mixtures may adopt an isotropic phase or form an aligned phase in magnetic fields. Well-resolved (1)H NMR spectra are observed in the isotropic or so-called fast-tumbling bicelle phase, over the range of temperatures investigated (10-40 degrees C), for molar ratios of long-chain lipid to short-chain lipid between 0.20 and 1.0. Small angle neutron scattering data of this phase are consistent with the model in which bicelles were proposed to be disk-shaped. The experimentally determined dimensions are roughly consistent with the predictions of R.R. Vold and R.S. Prosser (J. Magn. Reson. B 113 (1996)). Differential paramagnetic shifts of head group resonances of dimyristoylphosphatidylcholine (DMPC) and dihexanoylphosphatidylcholine (DHPC), induced by the addition of Eu(3+), are also consistent with the bicelle model in which DHPC is believed to be primarily sequestered to bicelle rims. Selective irradiation of the DHPC aliphatic methyl resonances results in no detectable magnetization transfer to the corresponding DMPC methyl resonances (and vice versa) in bicelles, which also suggests that DHPC and DMPC are largely sequestered in the bicelle. Finally, (1)H spectra of the antibacterial peptide indolicidin (ILPWKWPWWPWRR-NH(2)) are compared, in a DPC micellar phase and the above fast-tumbling bicellar phases for a variety of compositions. The spectra exhibit adequate resolution and improved dispersion of amide and aromatic resonances in certain bicelle mixtures.

Antimicrobial Cationic Peptides↗

Anomalous swelling in phospholipid bilayers is not coupled to the formation of a ripple phase.

Aligned stacks of monomethyl and dimethyl dimyristoyl phosphatidylethanolamine (DMPE) lipid bilayers, like the much studied dimyristoyl PC (DMPC) bilayers, swell anomalously in a critical fashion as the temperature is decreased within the fluid phase towards the main transition temperature, T(M). Unlike DMPC bilayers, both monomethyl and dimethyl DMPE undergo transitions into a gel phase rather than a rippled phase below T(M). Although it is not fully understood why there is anomalous swelling, our present results should facilitate theory by showing that the formation of the phase below T(M) is not related to critical phenomena above T(M).

Dimyristoylphosphatidylcholine↗

Method for obtaining structure and interactions from oriented lipid bilayers.

Precise calculations are made of the scattering intensity I(q) from an oriented stack of lipid bilayers using a realistic model of fluctuations. The quantities of interest include the bilayer bending modulus Kc, the interbilayer interaction modulus B, and bilayer structure through the form factor F(qz). It is shown how Kc and B may be obtained from data at large q(z) where fluctuations dominate. Good estimates of F(qz) can be made over wide ranges of q(z) by using I(q) in q regions away from the peaks and for q(r) not equal0 where details of the scattering domains play little role. Rough estimates of domain sizes can also be made from smaller q(z) data. Results are presented for data taken on fully hydrated, oriented DOPC bilayers in the L(alpha) phase. These results illustrate the advantages of oriented samples compared to powder samples.

Lipid Bilayers↗

Oblique membrane insertion of viral fusion peptide probed by neutron diffraction.

Fusion peptides mimic the membrane fusion activities of the larger viral proteins from which they derive their sequences. A possible mode of activity involves their oblique insertion into lipid bilayers, causing membrane disruption by promoting highly curved hemifusion intermediates, leading to fusion. We have determined the location and orientation of the simian immunodeficiency virus (SIV) fusion peptide in planar lipid bilayers using neutron lamellar diffraction. The helical axis of the peptide adopts an angle of 55 degrees relative to the membrane normal, while it positions itself nearest the lipid bilayer surface. This is the first direct observation of the structural interaction between a fusion peptide and a phospholipid bilayer.

Deuterium↗

Critical swelling in single phospholipid bilayers.

We approach the controversial anomalous swelling problem in membrane systems using small angle neutron scattering to measure relative changes in the bilayer thickness of unilamellar vesicles of dimyristoylphosphatidylcholine lipid bilayers in the vicinity of the main transition. These measurements conclusively demonstrate that at least half of the anomalous swelling previously observed in multilamellar vesicles of this system can be accounted for by the critical thickening of the bilayer itself, in contrast to conclusions drawn from several recent studies.

Dimyristoylphosphatidylcholine↗

Clarification of the ripple phase of lecithin bilayers using fully hydrated, aligned samples.

Aligned samples of lipid bilayers have been fully hydrated from water vapor in a different type of x-ray chamber. Our use of aligned samples resolves issues concerning the ripple phase that were ambiguous from previous powder studies. In particular, our x-ray diffraction data conclusively demonstrate that, on cooling from the L alpha to the P beta' phase, both chiral and racemic samples of dipalmitoyl phosphatidylcholine (DPPC) exhibit phase coexistence of long and short ripples with a ripple wavelength ratio lambda L/lambda S approximately 1.8. Moreover, the long ripple always forms an orthorhombic unit cell (gamma L = 90 degrees), strongly supporting the possibility that these ripples are symmetric. In contrast, gamma S for short ripples was consistently different from 90 degrees, implying asymmetric ripples. We continue to find no evidence that chirality affects the structure of rippled bilayers. The relative thermodynamic stability of the two types of ripples was investigated and a qualitative free energy diagram is given in which the long ripple phase is metastable. Finally, we suggest a kinetic mechanism, involving loss of water, that promotes formation of the metastable long ripple phase for special thermal protocols.

1,2-Dipalmitoylphosphatidylcholine↗