Biomedical subjects

R N Lewis

Publications and source records attributed to R N Lewis.

At least 37 records · Page 2Linked to original sources

PubMed1999

Calorimetric and spectroscopic studies of the effects of cholesterol on the thermotropic phase behavior and organization of a homologous series of linear saturated phosphatidylethanolamine bilayers.

Aqueous dispersions of cholesterol-containing phosphatidylethanolamine (PE) bilayers were examined by a combination of high-sensitivity differential scanning calorimetry (DSC), Fourier transform infrared (FTIR) and 31P-nuclear magnetic resonance spectroscopy. Regardless of hydrocarbon chain length, the incorporation of low levels of cholesterol into these bilayers causes progressive reductions in the temperature, enthalpy and overall cooperativity of the lipid hydrocarbon chain-melting phase transition. Moreover, at low cholesterol levels, the heating and cooling thermograms observed for the cholesterol/PE binary mixtures are similar, indicating comparable levels of lateral miscibility of cholesterol with PE bilayers in the gel and liquid-crystalline states. However, at higher levels of cholesterol incorporation, marked differences between the heating and cooling thermograms are noted. Upon heating, complex multicomponent thermograms are observed in PE bilayers containing large amounts of cholesterol, and the temperature and overall enthalpy values increase discontinuously from the pattern of monotonic decrease observed at lower cholesterol levels. Moreover, these discontinuities begin to emerge at progressively lower cholesterol concentrations as PE hydrocarbon chain length increases. Upon cooling, a simpler pattern of thermotropic behavior is observed, and the measured temperature and enthalpy values continue to decrease monotonically with increases in cholesterol content. These results suggest that at higher concentrations cholesterol exhibits a decreased degree of lateral miscibility in the gel or crystalline as compared to the liquid-crystalline states of PE bilayers, particularly in the case of the longer-chain PEs. Our FTIR and 31P-nuclear magnetic resonance spectroscopic studies also show that the thermotropic events observed with mixtures of low cholesterol content are analogous to the gel/liquid-crystalline phase transitions exhibited by the pure PEs. However, lamellar crystalline phases readily form when mixtures of high cholesterol content are cooled to low temperatures. Moreover, these crystalline phases are spectroscopically indistinguishable from those formed by the pure PEs, indicating that cholesterol is excluded from such phases. Upon subsequent heating, the melting of these crystalline phases gives rise to the complex thermograms detected by DSC and to the discontinuities in the phase transition temperature and enthalpy noted above. This pattern of behavior differs markedly from that observed with the corresponding phosphatidylcholines (PCs), where comparable degrees of cholesterol miscibility are observed in the gel and liquid-crystalline states even at high cholesterol concentrations, and where cholesterol inhibits rather than facilitates the formation of lamellar crystalline phases. We also find that the presence of cholesterol does not result in the hydrophobic mismatch-dependent shifts in the phase transition temperature in PE bilayers previously observed in PC bilayers of varying thickness. We attribute these differences in the effects of cholesterol on phospholipid thermotropic phase behavior to stronger electrostatic and hydrogen bonding interactions at the surfaces of PE and compared to PC bilayers.

Calorimetry, Differential Scanning↗

PubMed1999

The use of combined suprascapular and circumflex (articular branches) nerve blocks in the management of chronic arthritis of the shoulder joint.

Sixteen patients suffering from rheumatoid or osteoarthritis of the shoulder joint were studied. All patients complained of pain and limitation of active movement of the shoulder joint. Combined neural blockade of the suprascapular nerve (SSNB) and articular branches of the circumflex nerve (ACNB) was carried out using 4 mL of 1% prilocaine and 4 mL of 6% aqueous phenol. Following this procedure, the mean value for pain intensity decreased by 69% (VASP 2.7) and for abduction, adduction and flexion increased by 36-67% over a mean time of 13 weeks. Functional external and internal rotation of the shoulder joint also increased after neural blockade. These findings were significant (P < 0.05). Further clinical evaluation of combined SSNB and ACNB in relation to previously reported methods of neural blockade of the shoulder joint is warranted using a randomized, controlled, comparative study. Conventional power calculations (80% power, 5% test) indicate that 17 patients per group would be necessary to detect one standard deviation (about 2 VASP) or 64 per group to detect a change of 0.5 standard deviations.

Aged↗

PubMed1998

Molecular organization and dynamics of 1-palmitoyl-2-oleoylphosphatidylcholine bilayers containing a transmembrane alpha-helical peptide.

The molecular organization and dynamics have been investigated in membranes consisting of 1-palmitoyl-2-oleoyl-l-alpha-phosphatidylcholine (POPC) and various ratios of a transmembrane alpha-helical peptide, Ac-K2L24K2-amide (L24), in order to gain insights into how the transmembrane portions of membrane proteins are mixed with phospholipids and organized in biological membranes. Particular attention was paid to membranes with high peptide concentrations. The molecular organization and dynamics were studied in the ps-to-micros regime using various spin-labeling techniques. Conventional ESR spectra as well as saturation-recovery curves measured in both the presence and the absence of molecular oxygen showed that phosphatidylcholine spin-labels detect the existence of a single homogeneous environment, indicating that both L24 and POPC are likely to be undergoing fast translational diffusion in L24-POPC membranes of up to 9 mol % peptide. Since 16-18 molecules of phosphatidylcholine are required to surround a transmembrane alpha-helical peptide [Morrow, M. R., Huschilt, J. C., and Davis, J. H. (1985) Biochemistry 24, 5396-5406], L24 must form L24-rich regions at a P/L ratio of 1/10 instantaneously. However, these results suggest that the lipid exchange rates among the bulk, boundary, and L24-rich regions are fast, and that the L24-rich regions must be forming and dispersing rapidly in a time scale shorter than 0.1 micros, the conventional ESR spin-label time scale and the electron spin-lattice relaxation time scale in the presence of molecular oxygen. Although this does not exclude the possibility of the formation of small, stable oligomers of L24, it is unlikely because L24 lacks features that would favor their formation. L24 (9 mol %) increases the hydrophobicity of the central part of the POPC membrane from the level of 1-decanol to that of pure hexane and also increases the hydrophobicity near the membrane surface from the level of 2-propanol to that of 1-decanol. The effect of 9 mol % L24 on the order parameter profile is similar to that of decreasing the temperature by approximately 8 degrees C between 10 and 55 degrees C. It is concluded that L24 is highly miscible in POPC membranes even at high concentrations in the membrane.

Binding Sites↗

PubMed1998

Occurrence of monoacyl-diglucosyl-diacyl-glycerol and monoacyl-bis-glycerophosphoryl-diglucosyl-diacyl-glycerol in membranes of Acholeplasma laidlawii strain B-PG9.

It is shown by thin-layer and high-performance liquid chromatography that the two membrane lipids monoacyl-diglucosyl-diacyl-glycerol (MADGlcDAG) and monoacyl-bis-glycerophosphoryl-diglucosyl-diacyl-glycerol are synthesized by Acholeplasma laidlawii strain B-PG9 when the cells are grown in two different growth media. The two lipids are also synthesized by A. laidlawii strain A-EF22 and their chemical structures have been determined previously by NMR spectroscopy. Since a reversed hexagonal phase is the only liquid-crystalline phase formed by MADGlcDAG, it is concluded that A. laidlawii strain B-PG9, in resemblance to strain A-EF22, synthesizes three membrane lipids that are able to form reversed nonlamellar phases. A comparison of the membrane lipids from the two strains shows that there is essentially one lipid from each strain that differs. However, both these lipids have common physico-chemical properties, namely the ability to form reversed nonlamellar phases. Finally, it is also shown that novel lipids may be synthesized by A. laidlawii through long-time adaptation to altered growth conditions.

Acholeplasma laidlawii↗

PubMed1997

Nonlamellar phases induced by the interaction of gramicidin S with lipid bilayers. A possible relationship to membrane-disrupting activity.

The interactions of the cyclic peptide gramicidin S (GS) with a variety of single-component lipid bilayers, and with membrane polar lipid extracts of Acholeplasma laidlawii B and Escherichia coli, were examined by differential scanning calorimetry (DSC), 31P-nuclear magnetic resonance (NMR) spectroscopy, and X-ray diffraction. The DSC data indicate that the effects of GS on the thermotropic phase behavior of phosphatidylcholine and phosphatidylethanolamine dispersions are compatible with those expected of peptides interacting primarily with the polar headgroup and/or the polar/apolar interfaces of lipid bilayers. These DSC studies also suggest that GS exhibits stronger interactions with the more fluid bilayers. For mixtures of GS with lipids such as phosphatidylcholine, phosphatidylserine, cardiolipin, and sphingomyelin, axially symmetric 31P-NMR powder patterns are observed throughout the entire temperature range examined (0-90 degrees C), and there is little evidence for significant destabilization of the lipid bilayer with respect to nonlamellar phases. With mixtures of GS with either phosphatidylethanolamine, phosphatidylglycerol, or a nonlamellar phase-forming phosphatidylcholine, axially symmetric 31P-NMR powder patterns are also observed at low temperatures. However, at high temperatures, an isotropic component is observed in their 31P-NMR spectra, and the relative intensity of this component increases significantly with temperature and with GS concentration. Once formed at high temperatures, this isotropic component exhibits a marked cooling hysteresis and in most cases disappears only when the sample is recooled to temperatures well below the lipid hydrocarbon chain-melting phase transition temperature. We also show that GS induces the formation of isotropic components in the 31P-NMR spectra of heterogeneous lipid mixtures such as occur in A. laidlawii B and E. coli membranes. These observations suggest that GS induces the formation of cubic or other three dimensionally ordered inverted nonlamellar phases when it interacts with some types of lipid bilayers, a suggestion strongly supported by our X-ray diffraction studies. Our results also suggest that the capacity of GS to induce the formation of such phases increases with the intrinsic nonlamellar phase-preferring tendencies of the lipids with which it interacts probably by producing localized increases in membrane monolayer curvature stress. The latter effect could be part of the mechanism through which this peptide exhibits its antimicrobial and hemolytic activities.

Calorimetry, Differential Scanning↗

PubMed1997

Calorimetric and spectroscopic studies of the thermotropic phase behavior of the n-saturated 1,2-diacylphosphatidylglycerols.

The polymorphic phase behavior of a homologous series of n-saturated 1,2-diacyl phosphatidylglycerols (PGs) was studied by differential scanning calorimetry and Fourier transform infrared and 31P-nuclear magnetic resonance spectroscopy. When dispersed in aqueous media under physiologically relevant conditions, these compounds exhibit two thermotropic phase transitions that are structurally equivalent to the well-characterized pretransitons and gel/liquid-crystalline phase transitions exhibited by bilayers of the corresponding 1,2-diacyl phosphatidylcholines. Furthermore, when incubated at low temperatures, their gel phases spontaneously transform into one or more solid-like phases that appear to be highly ordered, quasicrystalline bilayers that are probably partially dehydrated. The quasicrystalline structures, which form upon short-term, low-temperature annealing of these lipids, are meta-stable with respect to more stable structures, to which they eventually transform upon prolonged low-temperature incubation. The rates of formation of the quasicrystalline phases of the PGs generally tend to decrease as hydrocarbon chain length increases, and PGs whose hydrocarbon chains contain an odd number of carbon atoms tend to be slower than those of neighboring even-numbered homologs. The calorimetric data also indicate that the quasicrystalline phases of these compounds become progressively less stable relative to both their gel and liquid-crystalline phases as the length of the hydrocarbon chain increases and that they decompose either to the liquid-crystalline phase (short- and medium-chain compounds) or to the normal gel phase (long-chain compounds) upon heating. The spectroscopic data indicate that although there is odd-even alternation in the structures of the quasicrystalline phases formed upon short-term low-temperature incubation of these compounds, the structural features of the stable quasicrystalline phases eventually formed are all similar. Furthermore, the degree of hydration and the nature of hydrogen bonding interactions in the headgroup and interfacial regions of these PG bilayers differ significantly from that observed in all other phospholipid bilayers studied so far. We suggest that many of the properties of PG bilayers can be rationalized by postulating that the glycerol moiety of the polar headgroup is directly involved in shielding the negative charges at the surface of the bilayer by means of hydration-like hydrogen bonding interactions with the phosphate moiety.

Calorimetry, Differential Scanning↗

PubMed1997

Respiratory complications of bone cement insertion during total hip replacement under spinal anaesthesia.

A report is made of respiratory complications which occurred following the insertion of Palacos bone cement during Howse total hip replacement under spinal anaesthesia in a 78-year-old female patient. Persistent cough and tachypnoea developed 30 s after the insertion of bone cement into the acetabulum and femur and persisted for 2-3 min producing considerable passive movement of the pelvis and femur. The probable cause, disadvantages and prevention of these complications are discussed.

Aged↗

PubMed1997

Some studies of the complement system during total hip replacement using bone cement under general anaesthesia.

Fourteen patients who underwent Charnley or Howse total hip replacement using CMW bone cement were studied. Mean values of C4 and C3 determined before the induction of anaesthesia and insertion of bone cement and also after this procedure showed no significant difference and were within the normal range. Furthermore, no evidence of C3 breakdown products was found. It was therefore concluded that neither the general anaesthetic technique nor the insertion of bone cement activated the classical or alternative complement pathways and such a mechanism was not responsible for the hypotension which occurred shortly after the insertion of bone cement in 10 cases studied.

Aged↗

PubMed1996

Differential scanning calorimetric study of the interaction of cholesterol with the major lipids of the Acholeplasma laidlawii B membrane.

It has been proposed that the lower levels of exogenous cholesterol incorporation into the membranes of the sterol-non-requiring as compared to the sterol-requiring mycoplasmas may be due to the much higher glycolipid content of the former and to the reduced ability of glycolipids, as opposed to phospholipids, to incorporate sterols [Efrati et al. (1986) Arch. Biochem. Biophys. 248, 282-288]. In order to test this hypothesis, we have investigated the interaction of cholesterol with the major membrane glyco- and phospholipids of the sterol-non-requiring mycoplasma Acholeplasma laidlawii B, utilizing elaidic acid-homogenous membranes in order to obviate any differences in the nature of cholesterol-lipid interactions due to variations in the fatty acid composition of the different membrane components. Specifically, we have studied the effect of increasing quantities of cholesterol on the thermotropic phase behavior of aqueous dispersions of phosphatidylglycerol, diglucosyl diacylglycerol, and monoglucosyl diacylglycerol, as well as the total membrane polar lipids of this organism, using high-sensitivity differential scanning calorimetry. We find that cholesterol is highly miscible in both the lamellar gel and liquid-crystalline states of phosphatidylglycerol but exhibits limited miscibility in the two neutral glycolipids, particularly in their lamellar gel and crystalline states. We also demonstrate that cholesterol has a limited miscibility in both the lamellar gel and liquid-crystalline states of bilayers composed of the total A. laidlawii B membrane polar lipids. These results demonstrate that the nature of cholesterol-lipid interactions depends markedly on the structure of the glycerolipid polar headgroup and suggests that the incorporation of lower levels of cholesterol into the membranes of the sterol-non-requiring mycoplasmas may indeed be due, at least in part, to their high glycolipid contents. We also show that cholesterol stabilizes the lamellar liquid-crystalline phase of the monoglucosyl diacylglycerol relative to the inverted hexagonal phase at all sterol concentrations, in contrast to the effects of cholesterol on dielaidoylphosphatidylethanolamine, which destabilizes the lamellar liquid-crystalline phase at low concentrations.

Acholeplasma laidlawii↗

PubMed1996

The interfacial structure of phospholipid bilayers: differential scanning calorimetry and Fourier transform infrared spectroscopic studies of 1,2-dipalmitoyl-sn-glycero-3-phosphorylcholine and its dialkyl and acyl-alkyl analogs.

The thermotropic phase behavior of aqueous dispersions of dipalmitoylphosphatidylcholine (DPPC) and its 1,2-dialkyl, 1-acyl 2-alkyl and 1-alkyl 2-acyl analogs was examined by differential scanning calorimetry, and the organization of these molecules in those hydrated bilayers was studied by Fourier transform infrared spectroscopy. The calorimetric data indicate that substitution of either or both of the acyl chains of DPPC with the corresponding ether-linked hydrocarbon chain results in relatively small increases in the temperature (< 4 degrees C) and enthalpy (< 1 kcal/mol) of the lipid chain-melting phase transition. The spectroscopic data reveal that replacement of one or both of the ester-linked hydrocarbon chains of DPPC with its ether-linked analog causes structural changes in the bilayer assembly, which result in an increase in the polarity of the local environments of the phosphate headgroups and of the ester carbonyl groups at the bilayer polar/apolar interface. The latter observation is unexpected, given that ester linkages are considered to be intrinsically more polar that ether linkages. This finding cannot be satisfactorily rationalized unless the conformation of the glycerol backbones of the analogs containing ether-linked hydrocarbon chains differs significantly from that of diacyl glycerolipids such as DPPC. A comparison of the alpha-methylene scissoring bands and the methylene wagging band progressions of these lipids with the corresponding absorption bands of specifically chain-perdeuterated analogs of DPPC also supports the conclusion that replacement of the ester-linked hydrocarbon chains of DPPC with the corresponding ether-linked analog induces conformational changes in the lipid glycerol backbone. The suggestion that the conformation of glycerol backbones in the alkyl-acyl and dialkyl derivatives of DPPC differs from that of the naturally occurring 1,2-diacyl glycerolipid suggests that mono- and di-alkyl glycerolipids may not be good models of their diacyl analogs. These results, and previously published evidence that DPPC analogs with ether-linked hydrocarbon chains spontaneously form chain-interdigitated gel phases at low temperatures, clearly indicate that the properties of lipid bilayers can be substantially altered by small changes in the chemical structures of their polar/polar interfaces, and highlight the critical role of the interfacial region as a determinant of the structure and organization of lipid assemblies.

1,2-Dipalmitoylphosphatidylcholine↗

PubMed1995

Quantitation of the phase preferences of the major lipids of the Acholeplasma laidlawii B membrane.

We have quantitated the phase preferences of all of the quantitatively significant lipids of fatty acid-homogeneous Acholeplasma laidlawii membranes by determining the effect of small amounts of each lipid on the lamellar/reversed hexagonal phase transition temperature of a phosphatidylethanolamine matrix of identical fatty acid composition using differential scanning calorimetry. We find that the incorporation of small amounts of these lipids produce effects ranging from a moderate depression to a marked elevation of the lamellar/reversed hexagonal phase transition temperature of the corresponding phosphatidylethanolamine. Thus, although the total membrane lipids from this organism form only lamellar phases under physiological conditions, the individual membrane lipids appear to exhibit a wide range of phase preferences. Phosphatidylglycerol and diglucosyldiacylglycerol seem to have relatively strong and weak preferences for the lamellar liquid-crystalline phase, respectively, while monoglucosyldiacylglycerol, and especially acyl polyprenyl glucoside, strongly prefers the reversed hexagonal phase. Most notable in this regard is the phase preference of glycerylphosphoryldiglucosyldiacylglycerol, which strongly destabilizes the reversed hexagonal phase and which we show in the accompanying paper [Lewis, R. N. A. H., & McElhaney, R. N. (1995) Biochemistry 34, 13818-13824] actually prefers the normal micellar phase in isolation. The presence of normal, lamellar, and reversed phase-preferring lipids in a single membrane has important implications for understanding the physical basis of lipid organization and biosynthetic regulation in this and possibly in other organisms. We also show that the characteristic effect of the individual A. laidlawii membrane lipids on the lamellar/reversed hexagonal phase transition temperature of the phosphatidylethanolamine matrix is not well correlated with their polar headgroup intrinsic volumes.(ABSTRACT TRUNCATED AT 250 WORDS)

Acholeplasma laidlawii↗

PubMed1995

Acholeplasma laidlawii B membranes contain a lipid (glycerylphosphoryldiglucosyldiacylglycerol) which forms micelles rather than lamellar or reversed phases when dispersed in water.

It has been proposed that each of the lipids from the Acholeplasma laidlawii membrane prefers to form either a lamellar or a reversed cubic or hexagonal phase when dispersed in excess water at physiologically relevant temperatures and ionic strengths. In this study, we have reinvestigated the thermotropic phase behavior of all the major membrane lipids from A. laidlawii B membranes derived from cells grown in equimolar palmitic and elaidic acids. We confirm that phosphatidylglycerol (PG) and diglucosyldiacylglycerol (DGDG) from such membranes do indeed form only lamellar phases over the temperature range 5-80 degrees C. We also confirm that the monoglucosyldiacylglycerol (MGDG) and acyl polyprenyl glucoside (APG) exist in lamellar phases at lower temperatures but do form reversed phases at higher temperatures. However, we present here optical, differential scanning calorimetric, quasielastic light scattering, and 2H- and 31P-nuclear magnetic resonance spectroscopic evidence indicating that one lipid component of the A. laidlawii B membrane, namely, glycerylphosphoryldiglucosyldiacylglycerol (GPDGDG), actually forms normal micellar rather than lamellar or reversed phases when dispersed in excess water at physiological temperatures. To the best of our knowledge, this is the first demonstration of the existence of a micellar phase-preferring lipid in a prokaryotic cell membrane, and only the second demonstration of the existence of a micellar phase-forming lipid in any biological membrane. We also show that GPDGDG levels change greatly depending on the fatty acid composition of the membrane lipids of this organism.(ABSTRACT TRUNCATED AT 250 WORDS)

Acholeplasma laidlawii↗

PubMed1995

Peptide models of helical hydrophobic transmembrane segments of membrane proteins. 1. Studies of the conformation, intrabilayer orientation, and amide hydrogen exchangeability of Ac-K2-(LA)12-K2-amide.

The secondary structure, amide hydrogen exchangeability, and intramembrane orientation of the hydrophobic peptide Ac-K2-(LA)12-K2-amide [(LA)12] were studied by a combination of circular dichroism (CD), Fourier transform infrared (FTIR), and proton nuclear magnetic resonance (1H NMR) spectroscopic techniques. All three techniques indicate that (LA)12 adopts predominantly helical conformations in various organic solvents, detergent micelles, and phospholipid bilayers. Also, attenuated total reflectance FTIR studies of oriented phospholipid bilayers demonstrate that (LA)12 is arranged with the long helical axis perpendicular to the bilayer plane. FTIR and 1H NMR studies of the exchangeability of the amide protons of (LA)12 indicate that in all media there are at least two populations of amide protons which exchange with the bulk solvent at markedly different rates. Moreover, the 1H NMR spectroscopic studies indicate that, in organic solvents and micellar dispersions, amide proton exchange rates decrease progressively from the N- or C-terminus of the peptide toward the central region. Our results are thus consistent with (LA)12 retaining a predominantly helical structure with so-called frayed ends in all media. The amide proton exchange studies also indicate that when (LA)12 is dispersed in lipid bilayers, the slowly exchanging population of amide protons is larger than that observed in organic solvents or in micellar dispersions and that most of that proton population is virtually unexchangeable. Such observations are consistent with the sequestration of the central regions of the peptide in the hydrophobic domains of the lipid bilayer. The CD and FTIR data indicate that although (LA)12 seems to retain conformations with a high alpha-helical content in all media examined, its conformation is sensitive to the composition of the surrounding medium, in contrast to the polyleucine-based analogues which have been studied previously. In particular, the FTIR spectroscopic data indicate that (LA)12 may exhibit an amide I absorption band between 1633 and 1639 cm-1 under some circumstances. The relative intensity of this band changes with the composition of the surrounding medium and its appearance has previously been correlated with the formation of 3(10)-helical structures [Miick et al. (1992) Nature 359, 653-655]. Thus (LA)12 may be interconverting between different helical conformations in response to changes in the physical properties of the medium in which the peptide is dispersed. Our results suggest that (LA)12 should serve as a good peptide model of hydrophobic, transmembrane helices which are conformationally sensitive to the properties of the lipid bilayer in which they reside.

Alanine↗

PubMed1995

Peptide models of helical hydrophobic transmembrane segments of membrane proteins. 2. Differential scanning calorimetric and FTIR spectroscopic studies of the interaction of Ac-K2-(LA)12-K2-amide with phosphatidylcholine bilayers.

The interactions of the hydrophobic helical transmembrane peptide Ac-K2-(LA)12-K2-amide [(LA)12] with a series of n-saturated diacylphosphatidylcholines (N:0 PC) were studied by high-sensitivity differential scanning calorimetry (DSC) and Fourier transform infrared (FTIR) spectroscopy. The incorporation of (LA)12 into these lipid bilayers results in a broadening of the chain-melting phase transitions of the lipids and progressive decreases in the characteristic temperatures and enthalpies of their gel/liquid-crystalline phase transitions. At low peptide/lipid ratios, the DSC thermograms exhibited by mixtures of (LA)12 with the short chain PCs (13:0 and 14:0) and with very long chain PCs (21:0 and 22:0) appear to be a summation of sharp and broad components, the former diminishing in intensity with increases in peptide concentration. This behavior can be approximated by that of a macroscopic mixture of peptide-poor and peptide-rich lipid domains, the relative proportions of which change with changes in peptide concentration. For peptide mixtures with the medium-chain PCs, the hydrocarbon chain-melting phase transition endotherms are not clearly resolvable into similar sharp and broad components. Instead, at all finite peptide concentrations the DSC heating thermograms appear as broad and highly asymmetric endotherms, the transition temperatures of which decrease significantly with increases in peptide concentration. For mixtures of (LA)12 with each of the lipids studied, the total hydrocarbon chain-melting transition enthalpy decreases with increasing peptide concentration but does not vanish at high peptide/lipid ratios. The FTIR spectra of (LA)12 in these PC bilayers indicate that the peptide retains a predominantly alpha-helical conformation in both the gel and liquid-crystalline phases of the short to medium chain PCs studied (N < 18). However, when incorporated into bilayers composed of the longer chain PCs (N > or = 18), (LA)12 undergoes a reversible conformational change at the gel/liquid-crystalline phase transition of the mixture. In the liquid-crystalline phase, the amide I regions of the FTIR spectra of these mixtures are indicative of a predominantly alpha-helical peptide conformation. However, upon freezing of the lipid hydrocarbon chains, populations and/or domains of (LA)12 giving rise to a sharp conformationally unassigned band near 1665 cm-1 are formed.(ABSTRACT TRUNCATED AT 400 WORDS)

Alanine↗

PubMed1995

The infrared dichroism of transmembrane helical polypeptides.

Polarized attenuated total internal reflectance techniques were applied to study the infrared dichroism of the amide I transition moment in two membrane-bound peptides that are known to form oriented transmembrane helices: gramicidin A in a supported phospholipid monolayer and Ac-Lys2-Leu24-Lys2-amide (L24) in oriented multibilayers. These studies were performed to test the ability of these techniques to determine the orientation of these peptides, to verify the value of optical parameters used to calculate electric field strengths, to examine the common assumptions regarding the amide I transition moment orientation, and to ascertain the effect of surface imperfections on molecular disorder. The two peptides exhibit marked differences in the shape and frequency of their amide I absorption bands. Yet both peptides are highly ordered and oriented with their helical axes perpendicular to the membrane surface. In the alpha-helix formed by L24, there is evidence for a mode with type E1 symmetry contributing to amide I, and the amide I transition moment must be more closely aligned with the peptide C=O (< 34 degrees) than earlier studies have suggested. These results indicate that long-standing assumptions about the orientation of amide I in a peptide require some revision, but that in general, infrared spectroscopy yields reliable information about the orientation of membrane-bound helical peptides.

1,2-Dipalmitoylphosphatidylcholine↗

PubMed1995

Interaction of a peptide model of a hydrophobic transmembrane alpha-helical segment of a membrane protein with phosphatidylethanolamine bilayers: differential scanning calorimetric and Fourier transform infrared spectroscopic studies.

High-sensitivity differential scanning calorimetry (DSC) and Fourier transform infrared (FTIR) spectroscopy were used to study the interaction of a synthetic alpha-helical hydrophobic transmembrane peptide, Acetyl-Lys2-Gly-Leu24-Lys2-Ala-Amide, and members of a homologous series of n-saturated diacylphosphatidylethanolamines (PEs). In the lower range of peptide mol fractions, the DSC endotherms exhibited by the lipid/peptide mixtures consist of two components. The temperature and cooperativity of the sharper, higher-temperature component are very similar to those of pure PE bilayers and are almost unaffected by variations in the peptide/lipid ratio. However, the fractional contribution of this component to the total enthalpy change decreases with increases in peptide concentration, and this component completely disappears at higher peptide mol fractions. The other component, which is less cooperative and occurs at a lower temperature, predominates at higher peptide concentrations. These two components of the DSC endotherm can be attributed to the chain-melting phase transitions of peptide-nonassociated and peptide-associated PE molecules, respectively. Although the temperature at which the peptide-associated PE molecules melt is progressively decreased by increases in peptide concentration, the magnitude of this shift is independent of the length of the PE hydrocarbon chain. In addition, the width of the phase transition observed at higher peptide concentrations is also relatively insensitive to PE hydrocarbon chain length, except that peptide gel-phase immiscibility occurs in very short- or very long-chain PE bilayers. Moreover, the enthalpy of the chain-melting transition of the peptide-associated PE does not decrease to 0 even at high peptide concentrations, suggesting that this peptide does not abolish the cooperative gel/liquid-crystalline phase transition of the lipids with which it is in contact. The FTIR spectroscopic data indicate that the peptide remains in a predominantly alpha-helical conformation, but that the peptide alpha-helix is subject to small distortions coincident with the changes in hydrophobic thickness that accompany the chain-melting phase transition of the PE bilayer. These data also indicate that the peptide significantly disorders the hydrocarbon chains of adjacent PE molecules in both the gel and liquid-crystalline states relatively independently of lipid hydrocarbon chain length. The relative independence of many aspects of PE-peptide interactions on the hydrophobic thickness of the host bilayer observed in the present study is in marked contrast to the results of our previous study of peptide-phosphatidylcholine (PC) model membranes (Zhang et al. (1992) Biochemistry 31:11579-11588), where strong hydrocarbon chain length-dependent effects were observed. The differing effects of peptide incorporation on PE and PC bilayers is ascribed to the much stronger lipid polar headgroup interactions in the former system. We postulate that the primary effect of transmembrane peptide incorporation into PE bilayers is the disruption of the relatively strong electrostatic and hydrogen-bonding interactions at the bilayer surface, and that this effect is sufficiently large to mask the effect of hydrophobic mismatch between the lengths of the hydrophobic core of the peptide and its host bilayer.

Amino Acid Sequence↗

PubMed1995

Relationship of body image and creative dance movement.

Findings supporting the proposition that dance movement improves a person's body image have been contradictory. Previous work focussed on styles such as ballet, jazz, and modern dance but it is arguable that creative dance movement, with its less structured approach and absence of predetermined performance standards, will have a positive influence on body image. This study examine scores on the Multidimensional Body-Self Relations Questionnaire of 112 women between 18 and 69 years who had been actively participating in creative dance movement courses for periods ranging from two weeks to 16.5 years. Subjects experienced in creative dance movement were more satisfied with their appearance, fitness, and body parts than subjects with less than five years of experience. Differences in evaluation of health were not clearly established. Given this analysis and the potential for wide-spread clinical use of creative dance movement with people having body-image disturbances, empirical research on the relationship between creative dance movement and body image is warranted.

Adolescent↗

PubMed1994

Studies of highly asymmetric mixed-chain diacyl phosphatidylcholines that form mixed-interdigitated gel phases: Fourier transform infrared and 2H NMR spectroscopic studies of hydrocarbon chain conformation and orientational order in the liquid-crystalline state.

Hydrocarbon chain conformational and orientational order in liquid-crystalline bilayers of the highly chain-asymmetric 1-O-eicosanoyl, 2-O-dodecanoyl and 1-O-decanoyl, 2-O-docosanoyl phosphatidylcholines were studied by Fourier transform infrared (FTIR) and deuterium nuclear magnetic resonance (2H-NMR) spectroscopy, respectively, and compared with appropriate symmetric-chain phosphatidylcholines at comparable reduced temperatures. FTIR spectroscopy indicates that these two asymmetric-chain phospholipids contain a slightly greater number of kink, a considerably larger number of double-gauche, but a somewhat smaller number of end-gauche conformers than does dipalmitoylphosphatidylcholine, a symmetric-chain phospholipid having the same total number of carbon atoms in its hydrocarbon chains. Moreover, the asymmetric-chain phospholipids also contain a larger total number of gauche conformers, suggesting that their hydrocarbon chains are more disordered overall than are those of dipalmitoylphosphatidylcholine. 2H-NMR studies of the specifically chain-perdeuterated analogs of these asymmetric-chain lipids reveal that the orientational order parameter profiles of their shorter and longer chains differ both qualitatively and quantitatively, regardless of whether they are esterified at the sn1- or sn2 positions of the glycerol molecule. The longer hydrocarbon chains exhibit unusual orientational order profiles in which the order gradient is steepest in the middle of the chain and relatively shallower in regions adjacent to the carboxyl and methyl termini, whereas the short hydrocarbon chains exhibit orientational order profiles typical of those commonly observed with conventional symmetric chain lipids. When compared at equivalent depths in the bilayer, the shorter hydrocarbon chains of the asymmetric-chain lipids are more orientationally disordered than are their longer chain counterparts. At comparable reduced temperatures, the shorter and longer chains of the asymmetric-chain lipids are more orientationally disordered than those of appropriate short and long symmetric-chain lipids, but the chain-averaged orientational order of the symmetric-chain lipid decreases more sharply with increases in temperature than does that of the comparable chain of the asymmetric-chain species. Moreover, the order plateau regions adjacent to the carboxyl groups of the longer chains of the asymmetric-chain phosphatidylcholines are shorter than those of symmetric-chain lipids of comparable hydrocarbon chain length. Overall, the data indicate that the conformational and orientational order in the liquid-crystalline states of these highly asymmetric-chain lipids differ significantly from those of comparable symmetric-chain lipids. Also, the unusual shape of the orientational order profile of the longer chains of the former is attributed to interaction between the methyl termini regions of the long chains with hydrocarbon chains in opposing monolayers. The latter suggests that some form of hydrocarbon chain interdigitation exists in liquid-crystalline bilayers of these highly asymmetric-chain lipids.

Deuterium↗