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1.
A critical step in the folding pathway of globular proteins is the formation of a tightly packed hydrophobic core. Several mutational studies have addressed the question of whether tight packing interactions are present during the rate-limiting step of folding. In some of these investigations, substituted side chains have been assumed to form native-like interactions in the transition state when the folding rates of mutant proteins correlate with their native-state stabilities. Alternatively, it has been argued that side chains participate in nonspecific hydrophobic collapse when the folding rates of mutant proteins correlate with side-chain hydrophobicity. In a reanalysis of published data, we have found that folding rates often correlate similarly well, or poorly, with both native-state stability and side-chain hydrophobicity, and it is therefore not possible to select an appropriate transition state model based on these one-parameter correlations. We show that this ambiguity can be resolved using a two-parameter model in which side chain burial and the formation of all other native-like interactions can occur asynchronously. Notably, the model agrees well with experimental data, even for positions where the one-parameter correlations are poor. We find that many side chains experience a previously unrecognized type of transition state environment in which specific, native-like interactions are formed, but hydrophobic burial dominates. Implications of these results to the design and analysis of protein folding studies are discussed. 相似文献
2.
The thermotropic properties and acyl chain packing characteristics of multilamellar dispersions of binary mixtures of 1-stearoyl-2-caprylphosphatidylcholine (C(18):C(10)PC), an asymmetric chain species, and dimyristoylphosphatidylcholine (C(14):C(14)PC), a symmetric chain lipid, were monitored by vibrational Raman spectroscopy. In order to examine each component of the binary mixture separately, the acyl chains of the symmetric chain species were perdeuterated. As shown by differential scanning calorimetry, the mismatch in the gel phase bilayer thickness between the two lipid components generates a lateral phase separation resulting in two distinct gel phases, G(I) and G(II), which coexist over much of the composition range. The Raman data demonstrate that the mixed interdigitated phase (three chains per headgroup), analogous to single component phase behavior, is retained when the C(18):C(10)PC component act as a host for the G(I) gel phase. In contrast, the C(18):C(10)PC molecules exhibit partial interdigitation (two chains per headgroup) when they are included as guests within the C(14):C(14)PC host matrix to form the G(II) gel phase. Compared to pure C(14):C(14)PC bilayers at equivalent reduced temperatures, the host G(II) gel phase C(14):C(14)PC molecules exhibit an increased acyl chain order, while for the host G(I) gel phase the C(14):C(14)PC lipid species show increased intrachain disorder. 相似文献
3.
Dependence of lipid chain and head group packing of the inverted hexagonal phase on hydration 总被引:2,自引:2,他引:0
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James R. Scherer 《Biophysical journal》1989,55(5):965-971
A model which positions the hydrophobic/hydrophilic boundary in phosphatidylethanolamine lipids at the first CH2 group in the acyl or alkyl chain is used to calculate the surface area per lipid, the mean chain and head-group dimensions and diameters of the hydrophilic tubes of the inverted hexagonal phase of didodecylphosphatidylethanolamine. The calculated surface areas compare favorably with areas obtained for the lamellar liquid crystal phase of the same lipid using the same boundary. Placement of the boundary within the lipid structure permits a determination of the maximum headgroup packing at hydration levels down to complete dehydration. The headgroup dimensions are consistent with a 5 Å diam void at the center of a hydrophilic tube at zero hydration. The calculated mean fluid chain length is ~2 Å smaller than the mean chain length of the lamellar phase at comparable levels of hydration. Comparison of the calculated mean fluid chain length and distances between hydrophobic boundaries shows that the fluid chains are interdigitated between adjacent tubes, and not interdigitated in the central space between three tubes. At low hydration the chains interdigitate in both spaces. The number of lipids packed around a tube at low hydration is only a function of the headgroup geometry, whereas at high hydration, it is a function of the number of carbon atoms in the chains. 相似文献
4.
Differences in hydrocarbon chain tilt between hydrated phosphatidylethanolamine and phosphatidylcholine bilayers. A molecular packing model. 总被引:16,自引:14,他引:2
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T J McIntosh 《Biophysical journal》1980,29(2):237-245
Both wide-angle and lamellar x-ray diffraction data are interpreted in terms of a difference in hydrocarbon chain tilt between fully hydrated dipalmitoyl phosphatidylcholine (DPPC) and dipalmitoyl phosphatidylethanolamine (DPPE). Although the hydrocarbon chains of multilayers of DPPC tilt ty approximately 30 degrees relative to the normal to the plane of the bilayer, as previously reported by others, the hydrocarbon chains of DPPE appear to be oriented approximately normal to the plane of the bilayer. It is found that the chain tilt in DPPC bilayers can be reduced by either: (a) adding an n-alkane to the bilayer interiors or (b) adding lanthanum ions to the fluid layers between bilayers. A molecular packing model is presented which accounts for these data. According to this model, DPPC chains tilt because of the size and conformation of the PC polar head group. 相似文献
5.
On the use of deuterium nuclear magnetic resonance as a probe of chain packing in lipid bilayers 总被引:3,自引:0,他引:3
The packing of hydrocarbon chains in the bilayers of lamellar (L alpha) phases of soap/water and phospholipid/water mixtures has been studied by deuterium NMR spectroscopy and X-ray diffraction. A universal correlation is shown to exist between the average C-D bond order parameter SCD of hydrocarbon chains and the average area per chain ach, irrespective of the chemical structure of the surfactant (hydrophilic group, number of chains per molecule, and chain length), composition, and temperature. The practical utility of the correlation is illustrated by its application to the characterization of the distribution of various hydrophobic and amphiphilic solutes in bilayers. The distribution of hydrocarbons within a bilayer is shown to depend upon their molecular structure in a manner which highlights the nature of the molecular interactions involved. For example, benzene is shown to be fairly uniformly distributed across the bilayer with an increasing tendency to distribute into the center at high concentrations. In contrast, the more complex hydrocarbon tetradecane preferentially distributes into the center of the bilayer at low concentrations, while at higher concentrations it intercalates between the surfactant chains. Alcohols such as benzyl alcohol, octanol, and decanol all interact similarly with the bilayer in so far as they are pinned to the polar/apolar interface, presumably by involvement of the hydroxyl group in a hydrogen bond. But the response of the surfactant chains to the void volume created in the center of the bilayer is dependent upon the distance of penetration of the alcohol into the bilayer. For benzyl alcohol, the shortest molecule, this void volume is taken up by the disordering of the chains, while for decanol, the longest molecule, it is absorbed by interdigitation of the chains of apposing monolayers. For octanol, the chain interdigitation mechanism is dominant at low concentrations, but there is a transition to chain disordering at high concentrations. Finally, it is shown that the correlation provides a useful test for statistical mechanical models of chain ordering in lipid bilayers. 相似文献
6.
Previous studies in this laboratory have delineated the relationship between the acyl chain asymmetry of mixed-chain phosphatidylcholines and the effect of ethanol concentration ([EtOH]) on their melting behavior (Li et al., Biophys J., 70 (1996) 2784-2794). This present investigation extends these findings to another phospholipid family by using high-resolution differential scanning calorimetry (DSC) to characterize the effect of ethanol concentration on the main phase transition temperature (Tm) of five molecular species of mixed-chain phosphatidylglycerol (PG). For C(14):C(18)PG, C(15):C(17)PG, C(16):C(16)PG, and C(17):C(15)PG, a biphasic profile in the Tm versus [EtOH] plot was observed, and the minimum in the plot for each PG occurred at 33, 15, 19, and 36 mg/ml, respectively. This biphasic behavior is typical of phospholipids whose acyl chain asymmetry is fairly small. For C(18):C(14)PG, only a linear decrease in the Tm was observed as a function of ethanol concentration; this effect is characteristic of highly asymmetric phospholipids. Our DSC results obtained with mixed-chain PG in the presence of ethanol demonstrate that the acyl chain asymmetry of the five lipids studied can be ranked as follows: C(15):C(17)PG相似文献
7.
The permeability transition pore (PTP) and the ATP-dependent potassium (mtK-ATP) channel of mitochondria are known to play key roles in mitochondrially mediated apoptosis. We investigated how modulation of the permeability transition pore (PTP) and the ATP-dependent potassium (mtK-ATP) channel, either as single elements or in combination, affects the proapoptotic intracellular calcium ([Ca(2+)](i)) transients and the mitochondrial membrane potential (psi(m)). For this purpose a model was established exploring the [Ca(2+)](i) transients in N2A cells using continuous application of ATP that causes a biphasic [Ca(2+)](i) response. This response was sensitive to endoplasmatic reticulum (ER) Ca(2+) depletion and a smooth ER Ca(2+)-ATPase (SERCA) antagonist. PTP inhibition by cyclosporine A (CsA) or its non-immunosuppressive derivative NIM811 caused an amplification of the secondary [Ca(2+)](i) peak and induced a hyperpolarization of psi(m). Both the putative mtK-ATP channel inhibitor 5-hydroxydecanoate (5-HD) and the opener diazoxide ameliorated the ATP-induced secondary [Ca(2+)](i) peak. The effect of diazoxide was accompanied by a depolarization of psi(m) whereas 5-HD had no effect on psi(m). When diazoxide and CsA or NIM811 were applied together the secondary [Ca(2+)](i) rise did not return to baseline and a not significant hyperpolarization of psi(m) was observed. So, simultaneous inhibition of PTP and activation of the mtK-ATP channel prevents the increased slope of the secondary [Ca(2+)](i) peak induced by CsA (or NIM811) and also the depolarization after diazoxide application. Hence, we propose that modulation of one of these channels leads to functional changes of the other channel by means of Delta[Ca(2+)](i) and Deltapsi(m). 相似文献
8.
《生物化学与生物物理学报:生物膜》2022,1864(1):183805
Continuous wave electron paramagnetic resonance spectroscopy of chain-labeled phospholipids is used to investigate the effects of hydration on the librational oscillations and the dynamical transition of phospholipid membranes in the low-temperature range 120–270 K. Bilayers of dipalmitoylphostatidiycholine (DPPC) spin-labeled at the first acyl chain segments and at the methyl ends and prepared at full, low, and very low hydration are considered. The segmental mean-square angular amplitudes of librations, 〈α2〉, are larger in the bilayer interior than at the polar/apolar interface and larger in the fully and low hydrated than in the very low hydrated membranes. For chain segments at the beginning of the hydrocarbon region, 〈α2〉-values are markedly restricted and temperature independent in DPPC with the lowest water content, whereas they increase with temperature in the low and fully hydrated bilayers, particularly at the highest temperatures. For chain segments at the chain termini, the librational amplitudes increase progressively, first slowly and then more rapidly with temperature in bilayers at any level of hydration. From the temperature dependence of the mean-square librational amplitude, the dynamical transition is detected around 240 K at the polar/apolar interface in fully and low hydrated DPPC and at around 225 K at the inner hydrocarbon region for bilayers at any hydration condition. At the dynamical transition the bilayers cross low energy barriers of activation energy in the range 10–20 kJ/mol. The results highlight biophysical properties of DPPC bilayers at low-temperature and provide evidence of the effects of the hydration on the dynamical transition in bilayers. 相似文献
9.
Direct visualization of asymmetric behavior in supported lipid bilayers at the gel-fluid phase transition
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We utilize in situ, temperature-dependent atomic force microscopy to examine the gel-fluid phase transition behavior in supported phospholipid bilayers constructed from 1,2-dimyristoyl-sn-glycero-3-phosphocholine, 1,2-dipentadecanoyl-sn-glycero-3-phosphocholine, and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine. The primary gel-fluid phase transition at T(m) occurs through development of anisotropic cracks in the gel phase, which develop into the fluid phase. At approximately 5 degrees C above T(m), atomic force microscopy studies reveal the presence of a secondary phase transition in all three bilayers studied. The secondary phase transition occurs as a consequence of decoupling between the two leaflets of the bilayer due to enhanced stabilization of the lower leaflet with either the support or the water entrained between the support and the bilayer. Addition of the transmembrane protein gramicidin A or construction of a highly defected gel phase results in elimination of this decoupling and removal of the secondary phase transition. 相似文献
10.
Progranulin haploinsufficiency causes biphasic social dominance abnormalities in the tube test
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A. E. Arrant A. J. Filiano B. A. Warmus A. M. Hall E. D. Roberson 《Genes, Brain & Behavior》2016,15(6):588-603
Loss‐of‐function mutations in progranulin (GRN) are a major autosomal dominant cause of frontotemporal dementia (FTD), a neurodegenerative disorder in which social behavior is disrupted. Progranulin‐insufficient mice, both Grn+/? and Grn ?/? , are used as models of FTD due to GRN mutations, with Grn+/? mice mimicking the progranulin haploinsufficiency of FTD patients with GRN mutations. Grn+/? mice have increased social dominance in the tube test at 6 months of age, although this phenotype has not been reported in Grn ?/? mice. In this study, we investigated how the tube test phenotype of progranulin‐insufficient mice changes with age, determined its robustness under several testing conditions, and explored the associated cellular mechanisms. We observed biphasic social dominance abnormalities in Grn+/? mice: at 6–8 months, Grn+/? mice were more dominant than wild‐type littermates, while after 9 months of age, Grn+/? mice were less dominant. In contrast, Grn ?/? mice did not exhibit abnormal social dominance, suggesting that progranulin haploinsufficiency has distinct effects from complete progranulin deficiency. The biphasic tube test phenotype of Grn+/? mice was associated with abnormal cellular signaling and neuronal morphology in the amygdala and prefrontal cortex. At 6–9 months, Grn+/? mice exhibited increased mTORC2/Akt signaling in the amygdala and enhanced dendritic arbors in the basomedial amygdala, and at 9–16 months Grn+/? mice exhibited diminished basal dendritic arbors in the prelimbic cortex. These data show a progressive change in tube test dominance in Grn+/? mice and highlight potential underlying mechanisms by which progranulin insufficiency may disrupt social behavior. 相似文献
11.
D L Reid S R Davidson T M Phernetton J H Rankin 《Journal of developmental physiology》1990,13(4):237-240
We have reported in a previous study that adenosine infusion causes fetal placental vascular resistance to increase after 2 min. To determine whether this action is followed by a more prolonged vasodilation, we studied 7 mature fetal lambs. At surgery, catheters were inserted into the fetal hindlimb arteries and veins. After a five day recovery period, control blood flow measurements were made by radiolabeled microsphere technique immediately after an infusion of 0.9% NaCl, (vehicle, 1.03 ml.min-1) into a fetal vein for 2 min. Within 5 min of the control blood flow measurement, adenosine (10 mg/min) was infused for 2 min. Blood flow measurements were repeated 5, 10, 15, 20 and 30 min after the end of the infusion period. Fetal arterial blood pressure dropped from 50 +/- 1 to 34 +/- 5 mmHg immediately after the adenosine infusion and returned to the control value within 5 min after the infusion. No further blood pressure response was detected. However, placental vascular resistance fell from 0.334 +/- 0.040 to 0.269 +/- 0.027 (P less than 0.05) at the 15 min measurement, remained low through the 20 min measurement (P less than 0.001) and was not different from control levels 30 min after the adenosine infusion. We conclude that the fetal placental vasculature responds to systemic adenosine infusion in a biphasic manner. The immediate reaction to adenosine is a transient vasoconstriction in the fetal placental vasculature followed by vasodilation 15 to 20 min after the initial exposure to adenosine. 相似文献
12.
Morphological behavior of lipid bilayers induced by melittin near the phase transition temperature
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Toraya S Nagao T Norisada K Tuzi S Saitô H Izumi S Naito A 《Biophysical journal》2005,89(5):3214-3222
Morphological changes of DMPC, DLPC, and DPPC bilayers containing melittin (lecithin/melittin molar ratio of 10:1) around the gel-to-liquid crystalline phase transition temperatures (Tc) were examined by a variety of biophysical methods. First, giant vesicles with the diameters of approximately 20 microm were observed by optical microscopy for melittin-DMPC bilayers at 27.9 degrees C. When the temperature was lowered to 24.9 degrees C (Tc = 23 degrees C for the neat DMPC bilayers), the surface of vesicles became blurred and dynamic pore formation was visible in the microscopic picture taken at different exposure times. Phase separation and association of melittin molecules in the bilayers were further detected by fluorescent microscopy and mass spectrometry, respectively. These vesicles disappeared completely at 22.9 degrees C. It was thus found that the melittin-lecithin bilayers reversibly undergo their fusion and disruption near the respective Tcs. The fluctuation of lipids is, therefore, responsible for the membrane fusion above the Tc, and the association of melittin molecules causes membrane fragmentation below the Tc. Subsequent magnetic alignments were observed by solid-state (31)P NMR spectra for the melittin-lecithin vesicles at a temperature above the respective Tcs. On the other hand, additional large amplitude motion induced by melittin at a temperature near the Tc breaks down the magnetic alignment. 相似文献
13.
In this communication we report the first systematic investigation of the thermodynamic properties of fully hydrated mixed-chain phosphatidylglycerols (PG) using high-resolution differential scanning calorimetry (DSC). The crystal structure of dimyristoylphosphatidylglycerol shows an acyl chain conformation that is nearly opposite to that of phosphatidylcholine (PC). In PC, the sn-1 chain is straight while the sn-2 chain contains a bend; for PG, the sn-1 contains a bend while the sn-2 chain is in the all-trans conformation (R.H. Pearson, I. Pascher, The molecular structure of lecithin dihydrate, Nature, 281 (1978) 499-501; I. Pascher, S. Sundell, K. Harlos, H. Eibl, Conformational and packing properties of membrane lipids: the crystal structure of sodium dimyristoylphosphatidylglycerol, Biochim. Biophys. Acta, 896 (1987) 77-88). If the structure of PG found in the single crystal can be extrapolated to that in the fully hydrated gel-state bilayer, the observed difference in acyl chain conformations implies that modulation of the acyl chain asymmetry will have an opposite effect on the thermotropic phase behavior of PG and PC. For example, it is expected, based on the crystal structures, that C(15):C(13)PG should have a higher main phase transition temperature (Tm) than C(14):C(14)PG, and C(13):C(15)PG should have a lower Tm than C(14):C(14)PG. However, our DSC studies show clearly that the expectation is not borne out by experimental data. Rather, the Tm values of C(15):C(13)PG, C(14):C(14)PG, and C(13):C(15)PG are 18.2 degrees C, 23.1 degrees C, and 24.4 degrees C, respectively. Several other PGs, each with a unique acyl chain composition, have also been studied in this laboratory using high-resolution DSC. It is shown that the acyl chain conformation of fully hydrated PG in general is nearly opposite to that seen in the PG crystal structure. 相似文献
14.
We show that the activity of an ion channel is correlated with the phase state of the lipid bilayer hosting the channel. By measuring unitary conductance, dwell times, and open probability of the K+ channel KcsA as a function of temperature in lipid bilayers composed of POPE and POPG in different relative proportions, we obtain that all those properties show a trend inversion when the bilayer is in the transition region between the liquid-disordered and the solid-ordered phase. These data suggest that the physical properties of the lipid bilayer influence ion channel activity likely via a fine-tuning of its conformations. In a more general interpretative framework, we suggest that other parameters such as pH, ionic strength, and the action of amphiphilic drugs can affect the physical behavior of the lipid bilayer in a fashion similar to temperature changes resulting in functional changes of transmembrane proteins. 相似文献
15.
Functional interactions of lipids and proteins were examined in brush-border membranes isolated from the kidney cortex by studying the temperature dependence of the hydrolytic enzyme activities. A close relationship was observed for the membrane proteins and the thermotropic lipid phase transitions. Three lines of evidences were provided for such dependence: a) Arrhenius relationship of the membrane-bound enzyme activities, and the effect of temperature in native and partially delipidated membranes, b) differential scanning calorimetric study of the membrane lipid phase transitions in the native and delipidated membranes, multilamellar vesicles prepared from the membrane extracted lipids, and in vesicles from dimyristoyl phosphatidylcholine, and c) the excimer (dimer)-formation studies of the membrane extrinsic fluorescent probe, pyrene, and the resultant membrane microviscosity. The brush-border membranes were partially delipidated with BuOH and 2,2,2-trifluoroethanol. The functional interactions of the delipidated membranes, which were greatly lost on lipid removal, were largely restored by the addition of exogenous lipids in the reconstitution process, which indicate the critical dependence of the membrane integral proteins on the neighboring lipid molecules in the bulk lipid phase. 相似文献
16.
Knowledge of the glass transition behavior of vitrification solutions is important for research and planning of the cryopreservation of biological materials by vitrification. This brief communication shows the analysis for the glass transition and glass stability of the multi-component vitrification solutions containing propanediol (PE), dimethyl sulfoxide (Me2SO) and polyvinyl alcohol (PVA) by using differential scanning calorimetry (DSC) during the cooling and subsequent warming between 25 and −150 °C. The glass formation of the solutions was enhanced by introduction of PVA. Partial glass formed during cooling and the fractions of free water in the partial glass matrix increased with the increasing of PVA concentration, which caused slight decline of glass transition temperature, Tg. Exothermic peaks of devitrification were delayed and broadened, which may result from the inhibition of ice nucleation or recrystallization of PVA. 相似文献
17.
《FEBS letters》2014,588(24):4776-4783
The central component AcrB of the Escherichia coli drug efflux complex AcrA–AcrB–TolC has been extensively investigated by X-ray crystallography of detergent–protein 3-D crystals. In these crystals, AcrB packs as trimers – the functional unit. We visualized the AcrB–AcrB interaction in its native environment by examining E. coli lipid reconstituted 2-D crystals, which were overwhelmingly formed by asymmetric trimers stabilized by strongly-interacting monomers from adjacent trimers. Most interestingly, we observed lattices formed by an arrangement of AcrB monomers distinct from that in traditional trimers. This hitherto unobserved packing, might play a role in the biogenesis of trimeric AcrB. 相似文献
18.
Mean-field calculations of chain packing and conformational statistics in lipid bilayers: comparison with experiments and molecular dynamics studies. 总被引:1,自引:1,他引:0
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A molecular, mean-field theory of chain packing statistics in aggregates of amphiphilic molecules is applied to calculate the conformational properties of the lipid chains comprising the hydrophobic cores of dipalmitoyl-phosphatidylcholine (DPPC), dioleoyl-phosphatidylcholine (DOPC), and palmitoyl-oleoyl-phosphatidylcholine (POPC) bilayers in their fluid state. The central quantity in this theory, the probability distribution of chain conformations, is evaluated by minimizing the free energy of the bilayer assuming only that the segment density within the hydrophobic region is uniform (liquidlike). Using this distribution we calculate chain conformational properties such as bond orientational order parameters and spatial distributions of the various chain segments. The lipid chains, both the saturated palmitoyl (-(CH2)14-CH3) and the unsaturated oleoyl (-(CH2)7-CH = CH-(CH2)7-CH3) chains are modeled using rotational isomeric state schemes. All possible chain conformations are enumerated and their statistical weights are determined by the self-consistency equations expressing the condition of uniform density. The hydrophobic core of the DPPC bilayer is treated as composed of single (palmitoyl) chain amphiphiles, i.e., the interactions between chains originating from the same lipid headgroup are assumed to be the same as those between chains belonging to different molecules. Similarly, the DOPC system is treated as a bilayer of oleoyl chains. The POPC bilayer is modeled as an equimolar mixture of palmitoyl and oleoyl chains. Bond orientational order parameter profiles, and segment spatial distributions are calculated for the three systems above, for several values of the bilayer thickness (or, equivalently, average area/headgroup) chosen, where possible, so as to allow for comparisons with available experimental data and/or molecular dynamics simulations. In most cases the agreement between the mean-field calculations, which are relatively easy to perform, and the experimental and simulation data is very good, supporting their use as an efficient tool for analyzing a variety of systems subject to varying conditions (e.g., bilayers of different compositions or thicknesses at different temperatures). 相似文献
19.
The main gel-fluid phase transition of wet lipid bilayers is examined in terms of a microscopic interaction model which incorporates both trans-gauche isomerism of the lipid acyl chains and crystal orientation variables for the lipid molecules. The model gives two scenarios for the phase behavior of wet lipid bilayers in terms of temperature: (i) chain melting occurs at a higher temperature than crystallization, or (ii) chain melting and crystallization occur at the same temperature. Experimental data for lipid bilayers is consistent with the second scenario. In this case, computer simulation is used to investigate the non-equilibrium behaviour of the model. The numerical data is intepreted in terms of interfacial melting on heating and grain formation on cooling through the main phase transition. Interfacial melting is a non-equilibrium process in which the grains of a polycrystalline bilayer melt inwards from the boundaries. The prediction of interfacial melting in wet lipid bilayers is examined in relation to data from both equilibrium and nonequilibrium measurements, to corresponding phase behavior in monolayers, and to previous theoretical work.Abbreviations DHPE
dihexadecyl phosphatidylethanolamine
- DMPA
dimyristoyl phosphatidic acid
- DMPC
dimyristoyl phosphatidylcholine
- DPPC
dipalmitoyl phosphatidylcholine
- DSC
differential scanning calorimetry
- MCS/S
Monte Carlo steps per site
Supported in part by the NSERC of Canada and FCAC du QuébecSupported by the Danish Natural Science Research Council under grant J.nr. 5.21.99.72 相似文献
20.
As one of its primary physiological functions, sPLA2-IIA appears to act as an antibacterial agent. In particular, sPLA2-IIA shows high activity towards Gram-positive bacteria such as Staphylococcus aureus (S. aureus). This antibacterial activity results from the preference of the enzyme towards membranes enriched in anionic lipids, which is a common feature of bacterial membranes. An intriguing aspect observed in a variety of bacterial membranes is the presence of a broad but cooperative lipid chain melting event where the lipids in the membrane transition from a solid-ordered (so) into a liquid-disordered (ld) state close to physiological temperatures. It is known that the enzyme is sensitive to the level of lipid packing, which changes sharply between the so and the ld states. Therefore, it would be expected that the enzyme activity is regulated by the bacterial membrane thermotropic behavior. We determine by FTIR the thermotropic lipid chain melting behavior of S. aureus and find that the activity of sPLA2-IIA drops sharply in the so state. The activity of the enzyme is also evaluated in terms of its effects on cell viability, showing that cell survival increases when the bacterial membrane is in the so state during enzyme exposure. These results point to a mechanism by which bacteria can develop increased resistance towards antibacterial agents that act on the membrane through a cooperative increase in the order of the lipid chains. These results show that the physical behavior of the bacterial membrane can play an important role in regulating physiological function in an in vivo system. 相似文献