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* Department of Physics and Astronomy, University of California, Irvine, California 92697-4575; Institute for Surface and Interface Science, University of California, Irvine, California 92697; and Department of Chemistry, University of California, Irvine, California 92697-2025
Correspondence: Address reprint requests to Douglas J. Tobias, Tel.: 949-824-4295; Fax: 949-824-8571; E-mail: dtobias{at}uci.edu.
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ABSTRACT |
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 TOP ABSTRACT INTRODUCTIONMETHODSRESULTS AND DISCUSSIONSUMMARYACKNOWLEDGEMENTSREFERENCES |
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INTRODUCTION |
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TOPABSTRACTINTRODUCTIONMETHODSRESULTS AND DISCUSSIONSUMMARYACKNOWLEDGEMENTSREFERENCES |
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90 wt % phospholipids and 10 wt % apoproteins, which is produced, secreted, and recycled by type II pneumocytes (Notter, 2000
). The main physiological action of pulmonary surfactant is the control of the surface tension during breathing, allowing the stability and proper mechanics of the alveoli.
Deficiency and dysfunction of pulmonary surfactant have been associated with several respiratory diseases (Frerking et al., 2001; Notter, 2000). The one with major incidence (Notter, 2000) is neonatal respiratory distress syndrome (NRDS) which affects premature infants. The conventional clinical therapy for neonatal respiratory distress syndrome is the replacement of the missing endogenous material, by delivering to the alveoli an exogenous surfactant mixture (Robertson and Halliday, 1998). The same concept has started to be applied as a therapy for other forms of lung disease (Frerking et al., 2001; Robertson and Halliday, 1998).
Both endogenous and exogenous pulmonary surfactant accomplish their function through synergistic effects between the physicochemical properties of their components. Although their main physiological action is the same, endogenous pulmonary surfactant has a complex and delicate life cycle that could account for the high diversity of components. On the other hand, a material useful for the replacement therapy is only required to perform the surface activity function, namely, reduction of the work of breathing, prevention of alveoli collapse, and promotion of lung expansion.
The notion of synergy between components arises from the competitive requirements of the physiological function. Essential to the surfactant function is the reduction of surface tension to values close to zero. This requires a close-packed, fairly ordered film, and accounts for the presence of dipalmitoyl phosphatidylcholine (DPPC) as a major component, 40 wt % in mammals (Veldhuizen et al., 1998). At the same time, to produce a uniform force throughout the lung during the entire breathing cycle, the mixture must be fluid enough to maintain its integrity over a large range of surface tension/alveolus area and have easy adsorption and respreading. This function is accomplished by the presence of unsaturated and anionic phospholipids and two of the four surfactant proteins (SP), SP-B and SP-C (Notter, 2000; Possmayer et al., 2001; Veldhuizen et al., 1998). SP-B and SP-C are small hydrophobic proteins of 78 residues and 35 residues, respectively (Haagsman and Diemel, 2001). Although it is still unclear whether or not SP-C has a specific function related to the film surface activity (Haagsman and Diemel, 2001; Possmayer et al., 2001), it has been shown by genetic ablation of the SP-B gene in mice (Clark et al., 1995), that SP-B is indispensable to the breathing function.
The comprehensive study by Tanaka et al. (Tanaka et al., 1986) showed that a mixture of DPPC, SP-B/C extracted from bovine lung surfactant, anionic phospholipids from natural extracts, and saturated fatty acids has the desired surface activity as described above. The most recent evidence from in vitro experiments supports the idea of synergistic effects in multicomponent lipid and lipid/protein mixtures. Captive bubble and Langmuir trough experiments (Discher et al., 1999; Piknova et al., 2001) have shown that films of a purified lipid mixture from extracts of calf pulmonary surfactant exist, over a large range of surface pressure up to 70 mN/m, as a biphasic mixture in which domains of a DPPC-rich liquid crystalline phase (liquid condensed or tilted phase) are surrounded by an isotropic liquid (liquid expanded) phase poor in DPPC. The action of SP-B and SP-C has been studied in Langmuir trough experiments on DPPC/DPPG (dipalmitoyl phosphatidylglycerol) (Krüger et al., 1999), DPPG/POPG (palmitoyl oleoyl phosphatidylglycerol) (Takamoto et al., 2001), and DPPC/POPG (Ding et al., 2001) systems, whereby it was demonstrated that the interaction between the anionic lipids and SP-B and/or SP-C promotes the integrity of a biphasic film over a large range of surface pressure/area and temperatures (Takamoto et al., 2001). In relation to the integrity of the film, a common outcome from these studies (Piknova et al., 2001; Zasadzinski et al., 2001) is that the retention of all the film components in the air/liquid interface region is related to a 2D to 3D transformation, apparently reversible, within the liquid isotropic phase.
Although the importance of the interaction between SP-B and anionic lipids to the proper performance of the physiological function seems to be clear, the mechanisms involved are not well understood (Possmayer et al., 2001; Zasadzinski et al., 2001). This is an aspect of primary importance to the development of new, more effective, and less expensive formulations for replacement therapy. The most successful replacement therapies are based on formulations that include animal pulmonary surfactant or pulmonary tissue extracts that contain the essential apoproteins SP-B and SP-C and are complemented with synthetic lipids (Notter, 2000). However, the risk of disease transmission, the desire for routine prophylactic applications, the need for an uniform composition, and the limited availability of the current formulations have prompted the development of synthetic alternatives that could address these demands while performing a similar function (Frerking et al., 2001; Johansson et al., 2001).
One approach to the design of new exogenous surfactant formulations is the combination of synthetic lipids with synthetic peptides based on amphipathic regions of the native sequence of human SP-B, or new sequences based on the principles for the surface activity function of SP-B (Bruni et al., 1991; Cochrane, 1998; Johansson et al., 2001; Longo et al., 1993). One of these synthetic peptides that has received considerable attention, and is the subject of the present study, is the SP-B N-terminal peptide SP-B1–25 (Bruni et al., 1991; Longo et al., 1993). SP-B1–25, an amphipathic peptide with sequence FPIPLPYCWLCRALIKRIQAMIPKG, has been shown to exhibit surface activity resembling that of the full SP-B protein (Lee et al., 1997; Takamoto et al., 2001). The intermittency of cationic residues and the presence of a hydrophobic region, in this and other SP-B related peptides, have been considered as structural motifs essential to the surface activity. Results of monolayer (Longo et al., 1993) and bilayer (Bruni et al., 1991) experiments with SP-B1–25 mutants suggested that the lipid/peptide interaction has both an electrostatic and hydrophobic character.
The surface activity and phase behavior of lipid monolayers with SP-B1–25 has been studied extensively (Bruni et al., 1991; Ding et al., 2001; Flanders et al., 2000; Flanders et al., 2002; Lee et al., 1997; Lee et al., 2001; Lipp et al., 1998; Lipp et al., 1997; Longo et al., 1993; Takamoto et al., 2001). In particular, a system that has received considerable attention is palmitic acid (PA)/SP-B1–25. PA is an important component in some of the formulations for replacement therapy (Notter, 2000), however, the relevance of these studies is primarily as a model system for the interaction of SP-B with anionic lipids.
Fatty acid monolayers have long been considered as suitable lipid monolayer model systems (Kaganer et al., 1999), due mainly to the great variety of phases they present as a consequence of a smaller headgroup than phospholipids. By controlling temperature and subphase composition their phase behavior is easily modified, and this has allowed a complete characterization at the microscopic and atomic scales (Kaganer et al., 1999).
In the case of PA, the gas-isotropic liquid-tilted condensed phase triple point occurs at room temperature (Lipp et al., 1998), and this has led to a comprehensive characterization of the SP-B1–25 peptide/lipid interaction effects on the monolayer constitution. Results from monolayer isotherms and fluorescence microscopy (Lee et al., 1997; Lipp et al., 1997) established that the SP-B N-terminal peptide SP-B1–25 mimics the behavior and effects of the full protein on lipid monolayers. Extensive characterization by diverse microscopy techniques (Flanders et al., 2000; Flanders et al., 2002; Lee et al., 1997; Lipp et al., 1997), grazing incidence x-ray diffraction (GIXD), and x-ray reflectivity (XRR) (Lee et al., 2001) have shown that as a result of interaction between SP-B1–25 and PA, a protein-rich fluid or disordered phase is formed in monophasic regions of the PA phase diagram where the tilted condensed or liquid crystalline phase and the untilted condensed (solidlike) phase are stable. The resulting films are biphasic with the disordered protein-rich phase surrounding the ordered PA condensed phase. This change from a single condensed phase in PA to a biphasic mixture in PA/SP-B and PA/SP-B1–25 has a dramatic effect on the stability of the monolayer, increasing its collapse surface pressure from 40 mN/m in pure PA to about and above 60 mN/m in the PA/SP-B1–25 and PA/SP-B systems, respectively (Lee et al., 1997; Lipp et al., 1997). The results of XRR revealed that SP-B1–25 is partially inserted in the monolayer disordered phase, forming and angle with the interface normal (Lee et al., 2001).
In this paper we report molecular dynamics simulations of the SP-B1–25 at 20 wt % in monolayers of PA with three different surface densities at the air-water interface. We have chosen this system for an initial atomic scale investigation of SP-B in lipid environments because its relatively small size and complexity make it tractable for all-atom modeling, and because it has been the subject of a series of experimental studies. We have validated the simulation initial model by a favorable comparison of our structural characterization with the results obtained from XRR measurements. We have developed a detailed description of the peptide location, orientation, conformation, and interactions, as well as the influence of the peptide on the conformation and packing of the lipid hydrocarbon chains, as a function of compression in the monolayer. The results presented here provide previously lacking microscopic detail that complements the large body of experimental data available on SP-B structure and function.
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METHODS |
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TOPABSTRACTINTRODUCTIONMETHODSRESULTS AND DISCUSSIONSUMMARYACKNOWLEDGEMENTSREFERENCES |
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) for the location of the peptide in the PA/SP-B1–25 system. In this model (Fig. 1), the peptide in a 
-helical conformation was inserted in the PA monolayer at an angle with respect to the normal to the air-water interface. The location is such that the side chains of the cationic residues Arg-12, Lys-16, and Arg-17 were located at the same level as the carboxyl headgroups, whereas residues 1–8 were accommodated in the region of the aliphatic chains.
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). In particular, the model reflects results of polarized FTIR spectroscopy that indicated an -helix conformation in residues 8–25 with an orientation of 45° to the lipid-water interface normal, electron spin resonance (ESR) data showing that a N-terminal spin probe was located near the interface and that the peptide was not aggregated in the lipid environment, and tryptophan fluorescence that suggested Trp-9 was associated with the lipid headgroups, and absent from the aqueous phase.
To directly compare with experimental results (Lee et al., 1997; Lee et al., 2001; Lipp et al., 1997), we have chosen to perform our study of the PA/SP-B1–25 system at 16°C. At this temperature PA monolayers are below their gas-liquid-tilted condensed phase triple point (Lee et al., 1997; Lipp et al., 1997). The monolayer condensation occurs through a gas to tilted condensed phase transformation and, after further compression, the system presents a second-order transition to an untilted condensed (solidlike) phase. As indicated in the previous section, the presence of the peptide affects the system phase behavior by inducing a disordered phase that persists up to the monolayer collapse. The results from GIXD and XRR (Lee et al., 2001) indicated that presence of the peptide does not affect the molecular packing of the untilted condensed phase. Moreover, although the isotherms exhibit a nonzero apparent compressibility in the region of the untilted condensed phase, due to the biphasic nature of the monolayer, the second-order transition point is still evident at high peptide concentrations.
We carried out simulations at three different values of area per lipid, corresponding to the following points of the 16°C isotherm for the PA 20-wt % SP-B1–25 system at pH 6.9 (0.15 M NaCl) (Lee et al., 1997; Lipp et al., 1997): the middle point (34 Å2/lipid), the final point (26 Å2/lipid) of the tilted condensed phase region, and the first point (24 Å2/lipid) of the untilted condensed phase region.
In all the simulations PA was taken to be fully ionized. The ionization state of a fatty acid monolayer depends on the pH and ionic strength of the aqueous subphase and the monolayer specific area. The ionization extent of a fatty acid as a function of these variables is unknown, however, estimates based on Gouy-Chapman theory are available (Lipp et al., 1997): for a pure water subphase the extent of ionization is negligible, with estimates in the range of 0.004–0.2%, and for a saline buffered subphase (pH 6.9, 150 mM NaCl) the values range from 24% at 17 Å2/molecule (16°C) to 39% at 40 Å2/molecule (28°C). Ideally, one should model a mixture of protonated and deprotonated fatty acids, and establish equilibrium of the lateral disposition of the species. However this is not plausible on the timescale that is presently feasible for all-atom lipid monolayer simulations. It is reasonable to assume that a peptide that carries a charge of +4 would prefer to interact with ionized lipids. Moreover, as indicated in the previous section, the electrostatic nature of the PA/SP-B1–25 interaction can be ascertained from the experimental evidence. We have therefore decided that, as a first attempt, it is reasonable to assume a completely ionized monolayer.
The basic simulation setup consisted of two monolayers of PA placed on opposite faces of a water slab. This configuration allows the study of two identical but independent systems in one single simulation. Each system was placed in the center of an orthorhombic cell (Table 1), with the longest direction normal to the air-water interface. A thickness of the water slab of 42 Å and a simulation cell height of 200 Å were selected in order to avoid long-range interactions along the direction normal to the air-water interface. This configuration allows the use of three-dimensional periodic boundary conditions for the simulation systems, as well as spherical boundary conditions for the long-range interactions.
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The CHARMM22 force field (MacKerell et al., 1998) was used for the lipids and peptides, and the TIP3P (Jorgensen et al., 1983) model was used for water. The smooth particle mesh Ewald method (Essman et al., 1995) was used to calculate the electrostatic interactions, and the van der Waals interactions and the real space part of the Ewald sum were cutoff at 10 Å using a spherical truncation scheme. The simulations were carried out at constant volume and a constant temperature of 16°C using Nosé-Hoover chain thermostats (Martyna et al., 1992). A reversible, multiple time step algorithm (Martyna et al., 1996) was used to integrate the equations of motion with a time step of 6 fs, and the lengths of bonds involving hydrogen atoms were held fixed using the SHAKE-RATTLE-ROLL algorithm (Martyna et al., 1996). The lengths of the simulations were 2.70 ns for the systems with specific areas 26 and 24 Å2/lipid, and 2.28 ns for the 34 Å2/lipid system. For all systems the analysis was carried out on the last 540 ps of the simulations.
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RESULTS AND DISCUSSION |
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) for their XRR results on a similar sample. Each profile is parsed into five regions: 1), water bulk 2), peptide-water 3), peptide-headgroup 4), peptide-tail, and 5), the remaining tail region. Box models for density profiles across interfaces are constructed by delimiting each region by a pair of step functions with opposite sign, which in turn, are convoluted with a Gaussian to account for overall roughness (Schalke and Lösche, 2000). Interfacial thermal broadening arises from two contributions: capillary waves and intrinsic thermal agitation. Molecular dynamics simulation systems like the ones reported here are, in general, not large enough to present capillary waves. Total electron density profiles only reflect the effect of volume exclusion, which is assumed to be of the same order as intrinsic thermal agitation. In general the effect of capillary waves accounts for 85% of the total roughness. Therefore in order to perform the least-squares fit of the total electron density profile to the proposed four-box model, we have taken as a fixed global roughness parameter the value of 2.3 Å reported by Lee et al (Lee et al., 2001). The corresponding fitted parameters are listed in Table 2.
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), the results from the simulations are within the same range, suggesting that the model presented here is a good candidate for the structural characterization of the PA/SP-B1–25 fluid mixture. A detailed match or quantitative comparison with the experimental results is precluded by the arbitrary choice of the global roughness, the possibility that the scattered intensity of specific peptide residues in the experiment is smeared out by intrinsic thermal fluctuations differing from the corresponding volume exclusion, and differences in ionization and hydration effects, given that the scattering experiments were conducted on pure water.
For the systems of higher density, 24 Å 2/lipid and 26 Å 2/lipid, the total thickness of the tail region is, respectively, 18.5 Å and 17.5 Å. Comparing these values with the one for an all-trans, upright pentadecanoic hydrocarbon chain of 18.765 Å, tilt angles of 9.6° and 22.0° are obtained. These values are in agreement with the results of direct calculations presented in Table 3, indicating that the chains are not perturbed globally by the presence of the peptide. It is reasonable to conclude that the 24 Å2/lipid and 26 Å2/lipid systems are at opposites sides of the second-order phase transition, in agreement with the isotherm reported by Lipp et al. (Lipp et al., 1997). The thickness values of the tail-peptide and head-peptide regions in the three simulated systems differ by less than 0.5 Å or half of the bin width. This suggests that the same portions of the peptide are accommodated in the monolayer hydrophobic and hydrophilic regions through the whole range of stability of the tilted condensed phase and up to the transition to the untilted condensed phase.
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of Lys-24, which appears to be in the water region; for 24 and 26 Å2/lipid the C of Arg-12 extends to the hydrophobic region. For all three systems the main peak of the distribution corresponds to the charged atoms of Lys-16 and Arg-17, which moves toward the center of the headgroup distribution as the system density increases. Changes in shape and displacement of the peptide distributions with specific area indicate that the peptide undergoes significant changes in its conformation, which are discussed in more detail below. In contrast, there are no significant changes in the location or shape of the lipid distributions that cannot be attributed to the normal ordering of the monolayer at the different specific areas. Both the water and the aliphatic chains distribution present asymmetric shapes produced by exclusion effects due to presence of the peptide. In the case of the density profile for the aliphatic chains, a nonuniform distribution is consistent with a local disruption of the orientational order as will be shown in detail in the next section.
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and Arg-17 C with headgroup oxygens and water oxygens. Representative results are shown in Fig. 4 for the 24-Å2/lipid system. The distribution functions for pairs formed with headgroup oxygens present a sharp first peak at 2.875 ± 0.25 Å for Lys-16 and 3.375 ± 0.25 Å for Arg-17. For all the systems under study, it was found that on average there are at least two different lipid headgroups in the first coordination shell of the charged molecular groups of Lys-16 and Arg-17. Detailed results are presented in Table 4. The average number of headgroups in contact with a charged residue increases with the density of the lipid monolayer, and the number of waters decreases. Although water oxygens were found in some cases in the first coordination shell, no correlation was found between residues and the amount of water present. Therefore, it does not seem relevant to specify a specific hydration level for these residues. Also, spatial correlations between the cationic residues and chlorine counterions were not found. These results suggest that electrostatic interactions between the peptide and solution counterions are unlikely.
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The model presented above is consistent with the behavior reported for SP-B and SP-B1–25 in the presence of anionic lipids. The specific interaction between bovine SP-B and DPPG headgroups in DPPC/DPPG bilayers was shown to induce ordering without considerably perturbing the interior of the membrane (Baatz et al., 1990). Krüger et al. (Krüger et al., 1999), employing SP-B/C in concentrations that closely resemble the endogenous lung surfactant and controlling the subphase pH, concluded that the interaction of the proteins with DPPC/DPPG monolayers had an electrostatic character. As was indicated in the previous section, all the experimental evidence points to a partial insertion of SP-B1–25 in PA and other lipid monolayers with anionic components through an electrostatic interaction.
Monolayer orientational order
As mentioned above, the presence of nonuniform distributions for the electron density of aliphatic chains (cf. Fig. 3) can be taken as an indication of local breaking of the lipid monolayer order. To gain insight into the different effects that the presence of the peptide could have on the monolayer ordering, we have conducted a detailed analysis of the aliphatic chains orientational order in the higher density systems. The analysis was performed separately for the two independent monolayers in each system.
The ordering of lipid monolayers in the condensed phases is strongly dependent on the aliphatic chain correlations. As a measure of the degree of chain packing and correlation, we have computed the skeletal order parameter (SCC) defined as
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j is the angle between the interface normal and the vector defined by the position of Cj-1 and Cj+1, and the angular brackets denote an average over molecules and time. The fully ordered all-trans untilted chains state corresponds to a value of SCC = 1, whereas a fully random orientation would give SCC = 0 (Ben-Shaul, 1995). We have complemented this analysis by calculating the average fraction of gauche conformations per bond. Fig. 5 shows plots of SCC versus carbon number for the 24-Å2/lipid and 26-Å2/lipid systems. The designation as upper and lower monolayer corresponds to the orientation shown in Fig. 1. Clearly, in these systems, there are two distinct levels of orientational order: for those chains far from the peptide a typical dependency of highly correlated packing is observed, whereas for those chains next to the peptide the orientational long range order appears to have been broken.
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The set of chains considered as being next to the peptide includes all of the peptide nearest neighbors and those second neighbors whose conformation appeared to be affected by the presence of the peptide (top view snapshots of each monolayer are presented in Fig. 7). They represent 25% and 23% of the total number of chains for 24 Å2/lipid and 26 Å2/lipid, respectively. The form of the SCC profile for these groups is consistent with a liquidlike isotropic phase (Ben-Shaul, 1995) where there is a monotonic decrease of the order parameter toward the terminal methyl group.
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The analysis of the monolayers with a specific area of 26 Å2/lipid reveals a similar behavior to the upper monolayer with 24 Å2/lipid. Nevertheless, a few significant difference were found: the SCC profiles for the chains near the peptide appear to follow closely the same trend as the profile for the ordered region (Fig. 5), and these systems present a more uniform distribution of gauche configurations along the chain (Fig. 6). These differences with respect to the system of higher density are readily attributable to the larger specific area.
Fluorescence microscopy experiments (Krüger et al., 1999; Lee et al., 1997) have shown that in lipid monolayer systems containing SP-B and SP-B1–25, the protein is located in the regions rich in the fluorescent-labeled lipid, suggesting that the lipid/SP-B mixture conforms to a fluid or disordered phase. At high surface pressure this fluidized phase forms a network around domains of ordered condensed phases. For the PA/SP-B1–25 system, contrasting their XRR and GIXD results, Lee et al. (Lee et al., 2001) concluded that the mixture PA/SP-B1–25 is disordered. Moreover, they modeled their PA/SP-B1–25 reflectivity spectra as a biphasic mixture of ordered PA and disordered PA/SP-B1–25 with a distribution of submicron sized domains. These experimental findings are consistent with the picture that arises from our analysis of the monolayer orientational order. Our results reveal that the action of the SP-B1–25 peptide on the order of the lipid monolayer has a local character. In other words, by taking advantage of a nonrestricted azimuthal orientation degree of freedom, a statistical picture of an untilted condensed phase can be consistent with having local conformational defects. This would allow the lipid/peptide mixture to coexist with ordered domains over a wide range of surface pressure, area fractions, and microstructural arrangements. The diverse forms in which disorder is introduced in the lipid monolayer condensed phases, manifested by the possibility of distributing gauche defects in either a localized way around the peptide or as small domains over larger areas, suggest that a mechanically stable boundary could exist between the peptide-rich fluid phase and the lipid ordered phase, allowing the formation of the patterns or textures that have been observed experimentally: a dispersion of the ordered condensed phase in a fluid phase matrix. This kind of microstructure appears to be required to perform the full breathing cycle continuously and reversibly.
Peptide orientation and conformation
As was shown before (cf. Figs. 2 and 3) the peptide remains firmly lodged in the monolayer for all the specific areas studied. However, its conformation changes as revealed by the changes in the density profiles shown in Fig. 3. These changes are consistent with the phase evolution of the lipid monolayer from a low density tilted condensed state at 34 Å2/lipid to a high density untilted condensed state at 24 Å2/lipid. Details of the peptide conformation as a function of the system specific area are shown in Fig. 8 and Table 3. In Fig. 8, the SP-B1–25 backbone dihedral angles (
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) from average structures (>540 ps) are plotted on Ramachandran maps. The values are plotted separately for each peptide in the independent monolayers in each simulation. The average angle between the peptide long axis (the largest moment of inertia) and the interface normal is listed for each system in Table 3. Taken together these results indicate that for the systems of higher density (24 and 26 Å2/lipid), as the specific area decreases, the corresponding decrease in the peptide tilt angle with respect to the interface normal is accompanied by a decrease in the fraction of right-handed -helical conformation. The divergence of the system at 34 Å2/lipid from this last result is an indication of a lack of conformational correlation between the lipid monolayer and the peptide.
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-helix segment occurs around Lys-16 and Arg-17, which the electron density profiles and pair distribution functions revealed as the core region for anchoring between the lipid monolayer and the peptide. The second form consists of two separate fractions extended along the monolayer hydrophobic and hydrophilic regions with Lys-16 and Arg-17 in a random conformation. These two conformations are correlated with the order in the monolayer: the single helix fraction corresponds to the upper monolayer with a more uniform distribution of gauche defects along the chains near the peptide, whereas the double partitioning corresponds to the lower monolayer whose gauche defects distribution is concentrated on both chain ends.
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The structural conformations of SP-B1–25 have been characterized in samples in organic solvents or phospholipid membrane mimetics (Gordon et al., 1996; Gordon et al., 2000). The most detailed analysis is found in a recent study that combines results of 13C-enhanced FTIR of SP-B1–25 in POPG liposomes with molecular modeling techniques (Gordon et al., 2000). These authors reported an -helix conformation for residues 8–22 and ß-sheet for residues 1–6. However, none of the experimental systems indicated could be considered as a model of pulmonary surfactant as they lack the necessary characteristics of high-density lipid packing and long-range orientational order. Moreover, it is conceivable that the peptide location and orientation in the lipid matrix could be different in lipid monolayers and bilayers. Consequently, structural arrangements in these systems do not manifest the lipid/peptide synergistic effects characteristic of the interaction in the pulmonary surfactant. In the context of these studies some attempts were made to qualitatively illustrate the location of SP-B1–25 in a lipid monolayer (Gordon et al., 1996; Lee et al., 2001). No comparison can be made between those representations and the results presented here, as the former do not constitute the outcome of a peptide modeling in an inhomogeneous lipid monolayer environment.
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SUMMARY |
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TOPABSTRACTINTRODUCTIONMETHODSRESULTS AND DISCUSSIONSUMMARYACKNOWLEDGEMENTSREFERENCES |
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ACKNOWLEDGEMENTS |
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TOPABSTRACTINTRODUCTIONMETHODSRESULTS AND DISCUSSIONSUMMARYACKNOWLEDGEMENTSREFERENCES |
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Submitted on June 19, 2002; accepted for publication October 31, 2002.
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REFERENCES |
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