RESUMEN
The lipid bilayer thickness d(L), the transbilayer distance of lipid phosphate groups d(pp/inf> and the lipid surface area A(L) of fluid hydrated bilayers of lamellar phases of egg phosphatidylcholine or dipalmitoylphosphatidylcholine containing N-alkyl-N,N-dimethylamine N-oxides (CnNO), 1,4-butanedi-ammonium-N,N'-dialkyl-N,N,N',N'-tetramethyl dibromides (GSn) or mono-hydrochlorides of [2-(alkyloxy)phenyl]-2-(1-piperidinyl)ethylesters of carbamic acid (CnA) were obtained by X-ray diffraction, and the bilayer thickness in extruded unilamellar dioleoylphosphatidylcholine vesicles containing C12NO was obtained by the neutron scattering. The values of d(L), d(pp/inf> and A(L) change linearly up to the 1:1 amphiphile:lipid molar ratio. The slopes of these dependencies increase for d(L) and d(pp/inf> and decrease for AL) with an increasing number of carbons n in the amphiphile long hydrocarbon substituent (18> or =n> or =8 for CnNO, 16> or =n> or =9 for GSn, 12> or =n> or =5 for CnA), while the opposite trends are observed for the short substituent (8> or =n>/=6 for CnNO, 9> or =n> or =7 for GSn, 5> or =n> or =3 for CnA). In case of long substituents, the effects on dL), dpp/inf> and AL) are caused by the decrease in the difference between the lipid and amphiphile hydrocarbon chain lengths and by the increase in their van der Waals attraction. The short substituent amphiphiles are mobile and exchange between multiple binding sites in the bilayer, minimizing the bilayer surface area.
Asunto(s)
Membrana Dobles de Lípidos/química , Neutrones , Dispersión de Radiación , Difracción de Rayos X , 1,2-Dipalmitoilfosfatidilcolina/química , Fenómenos Biofísicos , Biofisica , Relación Dosis-Respuesta a Droga , Liposomas/química , Espectroscopía de Resonancia Magnética , Fosfatidilcolinas/química , Tensoactivos/farmacología , TemperaturaRESUMEN
An unusual micelle was discovered in mixtures of the nonionic detergent octaethyleneglycol-mono-n-dodecylether with disaturated phospholipids such as 1,2-dimyristoyl-sn-glycero-3-phosphocholine or 1,2-dipalmitoyl-sn-glycero-3-phosphocholine in water. These mixtures undergo a structural transition upon cooling through the chain-melting temperatures of the respective phospholipids, resulting in the formation of mixed micelles. Structural features of the micellar particles were studied here by synchrotron x-ray scattering. The translucent micellar solutions showed characteristic wide-angle reflections that were attributed to ordered hydrocarbon chains, whereas the absence of small-angle x-ray reflections indicated that there is no long-range order in these mixtures. The presence of ordered phospholipid acyl chains was confirmed by differential scanning calorimetry and isothermal titration calorimetry. The endothermic differential scanning calorimetry signals observed in the up-scan mode were tentatively ascribed to chain melting and mixing of the components. Isothermal titration of the mixed-micellar solutions into an excess of the detergent octaethyleneglycol-mono-n-dodecylether resulted in sudden uptake of the latent heat by the gel-state phospholipids. The heat uptake per mol of phospholipid decreased with increasing detergent/phospholipid molar ratio. A simple geometric model is presented assuming that the dominating particle species in the mixtures is a discoidal phospholipid aggregate with ordered acyl chains, surrounded by a toroidal detergent hoop. The model implies that the fraction of ordered phospholipid chains decreases with increasing detergent/phospholipid molar ratio, in agreement with the calorimetric results and high-resolution NMR spectroscopy.
Asunto(s)
Detergentes/química , Micelas , Fosfolípidos/química , Rastreo Diferencial de Calorimetría , Cristalización , Espectroscopía de Resonancia Magnética , Dispersión de Radiación , TermodinámicaRESUMEN
A binary mixture of dimyristoylphosphatidylcholine (DMPC) and cholesterol displays a fluid miscibility gap under excess water conditions. Effects due to the imperfect miscibility of the two amphiphiles are studied near to and far from thermodynamic equilibrium by time-resolved small angle x-ray diffraction. The experiment discloses that this mixture phase separates when leaving the miscibility gap upon heating, a transition that is not included in current phase diagrams. This transition appears to be reversible and shows a temperature hysteresis of only a few degrees. We suggest a model in which the transition is driven with increasing temperature by a movement of the cholesterol away from the hydrophilic-hydrophobic interface toward the hydrophobic core of the bilayer.
Asunto(s)
Colesterol/química , Dimiristoilfosfatidilcolina/química , Fenómenos Biofísicos , Biofisica , Entropía , Modelos Estadísticos , Temperatura , Termodinámica , Factores de Tiempo , Difracción de Rayos X , Rayos XRESUMEN
Bundles of intact, tetanized skeletal muscle fibers from Rana temporaria were subjected to sinusoidal length oscillations in the frequency domain 100 Hz to 3 kHz while measuring force and sarcomere length. Simultaneously, intensity of the third-order x-ray reflection of the axial myosin unit cell (I(M3)) was measured using synchrotron radiation. At oscillation frequencies <1 kHz, I(M3) was distorted during the shortening phase of the sinusoid (i.e., where bundle length was less than rest length). Otherwise, during the stretch phase of oscillations at all frequencies, during the shortening phase of oscillations above 1 kHz, and for bundles in the rigor state, I(M3) was approximately sinusoidal in form. Mean I(M3) during oscillations was reduced by 20% compared to the isometric value, suggesting a possible change in S1 disposition during oscillations. However, the amplitude of length change required to produce distortion (estimated from the phase angle at which distortion was first evident) corresponded to that of a step release sufficient to reach the maximum I(M3), indicating a mean S1 disposition during oscillations close to that during an isometric tetanus. The mechanical properties of the bundle during oscillations were also consistent with an unaltered S1 disposition during oscillations.
Asunto(s)
Músculo Esquelético/química , Músculo Esquelético/fisiología , Rana temporaria , Actinas/química , Actinas/metabolismo , Animales , Fenómenos Biomecánicos , Simulación por Computador , Contracción Isométrica , Fibras Musculares Esqueléticas/química , Fibras Musculares Esqueléticas/fisiología , Miosinas/química , Miosinas/metabolismo , Sarcómeros/química , Sarcómeros/metabolismo , Relación Estructura-Actividad , Sincrotrones , Difracción de Rayos XRESUMEN
Formation of low-temperature ordered gel phases in several fully hydrated phosphatidylethanolamines (PEs) and phosphatidylcholines (PCs) with saturated chains as well as in dipalmitoylphosphatidylglycerol (DPPG) was observed by synchrotron x-ray diffraction, microcalorimetry, and densitometry. The diffraction patterns recorded during slow cooling show that the gel-phase chain reflection cooperatively splits into two reflections, signaling a transformation of the usual gel phase into a more ordered phase, with an orthorhombic chain packing (the Y-transition). This transition is associated with a small decrease (2-4 microl/g) or inflection of the partial specific volume. It is fully reversible with the temperature and displays in heating direction as a small (0.1-0.7 kcal/mol) endothermic event. We recorded a Y-transition in distearoyl PE, dipalmitoyl PE (DPPE), mono and dimethylated DPPE, distearoyl PC, dipalmitoyl PC, diC(15)PC, and DPPG. No such transition exists in dimyristoyl PE and dilauroyl PE where the gel L(beta) phase transforms directly into subgel L(c) phase, as well as in the unsaturated dielaidoyl PE. The PE and PC low-temperature phases denoted L(R1) and SGII, respectively, have different hydrocarbon chain packing. The SGII phase is with tilted chains, arranged in an orthorhombic lattice of two-nearest-neighbor type. Except for the PCs, it was also registered in ionized DPPG. In the L(R1) phase, the chains are perpendicular to the bilayer plane and arranged in an orthorhombic lattice of four-nearest-neighbor type. It was observed in PEs and in protonated DPPG. The L(R1) and SGII phases are metastable phases, which may only be formed by cooling the respective gel L(beta) and L(beta') phases, and not by heating the subgel L(c) phase. Whenever present, they appear to represent an indispensable intermediate step in the formation of the latter phase.
Asunto(s)
Geles/química , Fosfolípidos/química , 1,2-Dipalmitoilfosfatidilcolina/química , Rastreo Diferencial de Calorimetría , Densitometría , Hidrocarburos/química , Fosfatidilcolinas/química , Fosfatidiletanolaminas/química , Fosfatidilgliceroles/química , Protones , Temperatura , Factores de Tiempo , Difracción de Rayos XRESUMEN
Force, sarcomere length, and equatorial x-ray reflections (using synchrotron radiation) were studied in chemically skinned bundles of fibers from Rana temporaria sartorius muscle, activated by UV flash photolysis of a new photolabile calcium chelator, NP-EGTA. Experiments were performed with or without compression by 3% dextran at 4 degrees C. Isometric tension developed at a similar rate (t(1/2) = 40 +/- 5 ms) to the development of tetanic tension measured in other studies (Cecchi et al., 1991). Changes in intensity of equatorial reflections (I(11) t(1/2), 15-19 ms; I(10) t(1/2), 24-26 ms) led isometric tension development and were faster than for tetanus. During shortening at 0.14P(o), I(10) and I(11) changes were partially reversed (18% and 30%, respectively, compressed lattice), in agreement with intact cell data. In zero dextran, activation caused a compression of A-band lattice spacing by 0.7 nm. In 3% dextran, activation caused an expansion of 1.4 nm, consistent with an equilibrium spacing of 45 nm. But, in both cases, discharge of isometric tension by shortening caused a rapid lattice expansion of 1.0-1.1 nm, suggesting discharge of a compressive cross-bridge force, with or without compression by dextran, and the development of an additional expansive force during activation. In contrast to I(10) and I(11) data, these findings for lattice spacing did not resemble intact fiber data.
Asunto(s)
Ácido Egtácico/análogos & derivados , Contracción Muscular/fisiología , Músculo Esquelético/química , Músculo Esquelético/fisiología , Animales , Fenómenos Biofísicos , Biofisica , Calcio/metabolismo , Calcio/farmacología , Quelantes , Técnicas In Vitro , Contracción Muscular/efectos de los fármacos , Músculo Esquelético/efectos de los fármacos , Fotólisis , Rana temporaria , Sarcómeros/química , Sarcómeros/fisiología , Difracción de Rayos XRESUMEN
Increasing amounts of information emphasise the relevance of prevention, early diagnosis and early treatment of periodontal diseases in children. Children and adolescents affected by periodontal disease, mainly those who present a fast and severe attachment loss, are considered to be at risk of developing early or advanced periodontitis. Alternatively they may be presenting a reflex of systemic conditions affecting the periodontium. This study was aimed at verifying the acceptability of the use of the Simplified PSR (Periodontal Screening and Recording) Index in a very young population since a previous study in Bahia, Brazil, indicated a very high need for periodontal treatment in adolescents and young adults. A total of 200 children aged 3-6 years from private schools in Bahia, Brazil, were examined by four trained undergraduate students. The screening system was well accepted by the subjects and the fact that it is a fast tool was considered important for the successful examination of all sextants without behavioural disturbance. A high prevalence and a low severity of parameters related to periodontal disease were found in this population. A statistically higher prevalence of PSR code 2 (61.5%) [54.50-68.49] CI 95% when compared to PSR codes 0 (23.5%) 117.93-30.10] CI 95%, 1 (14.5%) 110.07-20.32] CI 95% and 3 (0.5%) 10.02-3.18] CI 95% were shown. There was no statistically significant difference between female and male children for any PSR code. The finding of more parameters related to periodontal health in S5 when compared to sextants S6 and S4 showed statistical significance (CI 95%).
Asunto(s)
Tamizaje Masivo/métodos , Enfermedades Periodontales/epidemiología , Brasil/epidemiología , Niño , Preescolar , Intervalos de Confianza , Cálculos Dentales/epidemiología , Femenino , Hemorragia Gingival/epidemiología , Humanos , Masculino , Enfermedades Periodontales/clasificación , Bolsa Periodontal/epidemiología , Proyectos Piloto , Prevalencia , Factores SexualesRESUMEN
The structure of native yeast tRNA(Phe) and wheat germ ribosomal 5S RNA induced by different magnesium ion concentrations was studied in solution with a synchrotron mediated hydroxyl radical RNA cleavage reaction. We showed that very small amounts of Mg+2 can induce significant changes in the hydroxyl radical cleavage pattern of tRNA(Phe). It also turned out that a reactivity of tRNAz(Phe) towards *OH coincides with the strong metal binding sites. Because of the Mg ions are heavily hydrated one can suggest the strong correlation of the observed nucleosides reactivity in vicinity of Mg2+ binding sites with availability of water molecules as a source of hydroxyl radical. On the other hand the structure of wheat germ 5S rRNA is less sensitive to the hydroxyl radical reaction than tRNA(Phe) although some changes are visible at 4 mM Mg ions. It is probably due to the lack of strong Mg+2 binding sites in that molecule. The reactivity of nucleotides in loops C and D of 5S rRNA is not effected, what suggests their flexibility or involvement in higher order structure formation. There is different effect of magnesium on tRNA and 5S rRNA folding. We found that nucleotides forming strong binding sites for magnesium are very sensitive to X-ray generated hydroxyl radical and can be mapped with *OH. The results show, that guanine nucleotides are preferentially hydrated. X-ray footprinting mediated hydroxyl radical RNA cleavage is a very powerful method and has been applied to studies of stable RNAs for the first time.
Asunto(s)
Magnesio/farmacología , ARN Ribosómico 5S/química , ARN Ribosómico 5S/efectos de los fármacos , ARN de Transferencia de Fenilalanina/química , ARN de Transferencia de Fenilalanina/efectos de los fármacos , Secuencia de Bases , Sitios de Unión , Radical Hidroxilo , Modelos Moleculares , Datos de Secuencia Molecular , Conformación de Ácido Nucleico/efectos de los fármacos , ARN de Hongos/química , ARN de Hongos/efectos de los fármacos , ARN de Hongos/genética , ARN de Planta/química , ARN de Planta/efectos de los fármacos , ARN de Planta/genética , ARN Ribosómico 5S/genética , ARN de Transferencia de Fenilalanina/genética , Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/genética , Sincrotrones , Triticum/química , Triticum/genéticaRESUMEN
Stereochemically pure archeal acyclic bola-amphiphilic diphosphates 4 and 5, with the basic structure of the phospholipids found in Sulfolobus, have been synthesized for the first time. The self-assembly properties have been compared with those of the nearly identical 72-membered macrocyclic tetraether phosphates 3a and 3b, analogues of the major phospholipid components of Sulfolobus, Thermoplasma, and methanogenic Archea, which were also synthesized. Phase contrast and fluorescence microscopies have shown that the dipolar lipids 1 and 2 spontaneously formed vesicles. Whereas the macrocyclic dipolar phosphates 3 spontaneously formed vesicles (phase contrast and fluorescence microscopies), the bolaform phosphate 4 gave only a lamellar structure (synchrotron diffraction pattern: repeat distance of about 4.25 nm but with only a few layers). However, upon addition of the unphosphorylated precursors phytanol, phytol, or geranylgeraniol to the acyclic lipids 4 and 5, giant vesicles were rapidly formed. Addition of n-hexadecanol or cholesterol did not lead to vesicle formation. Therefore it was concluded that this vesicle formation occurs only when the added molecule is closely compatible with the constituents of the lipid layer and can be inserted into the double layer. A slight mismatch (cholesterol or n-hexadecanol/polyprenyl chains) is therefore enough to block the insertion process presumably required for vesicle formation.
RESUMEN
The shape of free Thermus flavus 5 S rRNA in solution at 1.3 nm resolution is restored from synchrotron x-ray scattering data using an ab initio simulated annealing algorithm. The free 5 S rRNA is a bent elongated molecule displaying a compact central region and two projecting arms, similar to those of the tRNA. The atomic models of the 5 S rRNA domains A-D-E and B-C in the form of elongated helices can be well accommodated within the shape, yielding a tentative model of the structure of the free 5 S rRNA in solution. Its comparison with the recent protein-RNA map in the ribosome (Svergun, D. I., and Nierhaus, K. H. (2000) J. Biol. Chem. 275, 14432-14439) indicates that the 5 S rRNA becomes essentially more compact upon complex formation with specific ribosomal proteins. A conceivable conformational change involves rotation of the B-C domain toward the A-D-E domain. The model of free 5 S rRNA displays no interactions between domains E and C, but such interactions are possible in the bound molecule.
Asunto(s)
ARN Ribosómico 5S/química , Dispersión de Radiación , Thermus/química , Thermus/genética , Algoritmos , Relación Dosis-Respuesta a Droga , Escherichia coli/metabolismo , Modelos Moleculares , Conformación de Ácido Nucleico , Conformación Proteica , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína , Ribosomas/química , Programas Informáticos , Rayos XRESUMEN
Experimental evidence supporting the hypothesis of gel-liquid crystalline phase coexistence in the stable ripple phase of diacylphosphatidylcholines has been obtained from time-resolved X-ray small- (SAXS) and wide-angle diffraction (WAXS) in the millisecond to second time domain. The pretransition of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) exhibits a thin lamellar liquid crystalline intermediate phase (designated Lalpha) if driven far away from equilibrium by an infrared temperature jump (T-jump) technique. The findings can be described by a two-step model. (1) Instantaneously with the T-jump, an anomalously thin lamellar liquid crystalline intermediate phase (d = 5.6-5.8 nm) forms, coexisting with the original gel-phase Lbeta'. Within the first seconds, the lamellar repeat distance of the intermediate increases to a value of about 6.7 nm. A closer examination of these kinetics reveals two relaxation components: a fast process, proceeding within tenths of a second, and a slow process, on the time scale of a few seconds. (2) Finally, both the liquid crystalline and the gel-phase relax into the stable ripple phase Pbeta'. The total process time of the transition is nearly independent of the addition of NaCl, but varies strongly with the chain length of the lecithin species.
Asunto(s)
1,2-Dipalmitoilfosfatidilcolina/química , Cristalización , Geles , Cinética , Modelos Moleculares , Conformación Molecular , Termodinámica , Difracción de Rayos XRESUMEN
Incorporation of the helical antimicrobial peptide alamethicin from aqueous phase into hydrated phases of dioleoylphosphatidylethanolamine (DOPE) and dioleoylphosphatidylcholine (DOPC) was investigated within a range of peptide concentrations and temperatures by time-resolved synchrotron X-ray diffraction. It was found that alamethicin influences the organizations of the non-bilayer-forming (DOPE) and the bilayer-forming (DOPC) lipids in different ways. In DOPC, only the bilayer thickness was affected, while in DOPE new phases were induced. At low peptide concentrations (<1.10(-4) M), an inverted hexagonal (H(II)) phase was observed as with DOPE dispersions in pure buffer solution. A coexistence of two cubic structures was found at the critical peptide concentration for induction of new lipid/peptide phases. The first one Q224 (space group Pn3m) was identified within the entire temperature region studied (from 1 to 45 degrees C) and was found in coexistence with H(II)-phase domains. The second lipid/peptide cubic structure was present only at temperatures below 16 degrees C and its X-ray reflections were better fitted by a Q212 (P4(3)32) space group, rather than by the expected Q229 (Im3m) space group. At alamethicin concentrations of 1 mM and higher, a nonlamellar phase transition from a Q224 cubic phase into an H(II) phase was observed. Within the investigated range of peptide concentrations, lamellar structures of two different bilayer periods were established with the bilayer-forming lipid DOPC. They correspond to lipid domains of associated and nonassociated helical peptide. The obtained X-ray results suggest that the amphiphilic alamethicin molecules adsorb from the aqueous phase at the lipid head group/water interface of the DOPE and DOPC membranes. At sufficiently high (>1.10(-4) M) solution concentrations, the peptide is probably accommodated in the head group region of the lipids thus inducing structural features of mixed lipid/peptide phases.
Asunto(s)
Alameticina/química , Antibacterianos/química , Membrana Dobles de Lípidos/química , Secuencia de Aminoácidos , Sustancias Macromoleculares , Modelos Moleculares , Datos de Secuencia Molecular , Fosfatidilcolinas , Fosfatidiletanolaminas , Conformación Proteica , Termodinámica , Agua , Difracción de Rayos XRESUMEN
Ceramide has recently been established as a central messenger in the signaling cascades controlling cell behavior. Physicochemical studies have revealed a strong tendency of this lipid toward phase separation in mixtures with phosphatidylcholines. The thermal phase behavior and structure of fully hydrated binary membranes composed of dimyristoylphosphatidylcholine (DMPC) and N-palmitoyl-ceramide (C16:0-ceramide, up to a mole fraction X(cer) = 0.35) were resolved in further detail by high-sensitivity differential scanning calorimetry (DSC) and x-ray diffraction. Both methods reveal very strong hysteresis in the thermal phase behavior of ceramide-containing membranes. A partial phase diagram was constructed based on results from a combination of these two methods. DSC heating scans show that with increased X(cer) the pretransition temperature T(p) first increases, whereafter at X(cer) > 0.06 it can no longer be resolved. The main transition enthalpy DeltaH remains practically unaltered while its width increases significantly, and the upper phase boundary temperature of the mixture shifts to approximately 63 degrees C at X(cer) = 0.30. Upon cooling, profound phase separation is evident, and for all of the studied compositions there is an endotherm in the region close to the T(m) for DMPC. At X(cer) >/= 0.03 a second endotherm is evident at higher temperatures, starting at 32.1 degrees C and reaching 54.6 degrees C at X(cer) = 0.30. X-ray small-angle reflection heating scans reveal a lamellar phase within the temperature range of 15-60 degrees C, regardless of composition. The pretransition is observed up to X(cer) < 0.18, together with an increase in T(p). In the gel phase the lamellar repeat distance d increases from approximately 61 A at X(cer) = 0. 03, to 67 A at X(cer) = 0.35. In the fluid phase increasing X(cer) from 0.06 to 0.35 augments d from 61 A to 64 A. An L(beta')/L(alpha) (ripple/fluid) phase coexistence region is observed at high temperatures (from 31 to 56.5 degrees C) when X(cer) > 0.03. With cooling from temperatures above 50 degrees C we observe a slow increase in d as the coexistence region is entered. A sudden solidification into a metastable, modulated gel phase with high d values is observed for all compositions at approximately 24 degrees C. The anomalous swelling for up to X(cer) = 0.30 in the transition region is interpreted as an indication of bilayer softening and thermally reduced bending rigidity.
Asunto(s)
Ceramidas , Dimiristoilfosfatidilcolina/química , Liposomas/química , Esfingosina/análogos & derivados , Fenómenos Biofísicos , Biofisica , Rastreo Diferencial de Calorimetría , Fluidez de la Membrana , Dispersión de Radiación , Esfingosina/química , Rayos XRESUMEN
A clinical case of Langerhans' cell histiocytosis, type eosinophilic granuloma, in a young adult patient is presented. Because of the occurrence of oral manifestations in initial stages of the disease, there is a need for a differential diagnosis, especially with the early-onset periodontitis.
Asunto(s)
Granuloma Eosinófilo/complicaciones , Enfermedades Periodontales/etiología , Adulto , Pérdida de Hueso Alveolar/etiología , Diagnóstico Diferencial , Estudios de Seguimiento , Hemorragia Gingival/etiología , Recesión Gingival/etiología , Humanos , Masculino , Pérdida de la Inserción Periodontal/etiología , Bolsa Periodontal/etiología , Periodontitis/diagnóstico , Pérdida de Diente/etiología , Movilidad Dentaria/etiologíaRESUMEN
Equatorial X-ray diffraction patterns were recorded from small bundles of one to three chemically skinned frog sartorius muscle fibres (time resolution 250 microseconds) during rapid stretch and subsequent release. In the relaxed state, the dynamic A-band lattice spacing change as a result of a 2 % step stretch (determined from the positions of the 10 and 11 reflections) resulted in a 21 % increase in lattice volume, while static studies of spacing and sarcomere length indicated than an increase in volume of >/=50 % for the same length change. In rigor, stretch caused a lattice volume decrease which was reversed by a subsequent release. In activated fibres (pCa 4.5) exposed to 10 mM 2,3-butanedione 2-monoxime (BDM), stretch was accompanied by a lattice compression exceeding that of constant volume behaviour, but during tension recovery, compression was partially reversed to leave a net spacing change close to that observed in the relaxed fibre. In the relaxed state, spacing changes were correlated with the amplitude of the length step, while in rigor and BDM states, spacing changes correlated more closely with axial force. This behaviour is explicable in terms of two components of radial force, one due to structural constraints as seen in the relaxed state, and an additional component arising from cross-bridge formation. The ratio of axial to radial force for a single thick filament resulting from a length step was four in rigor and BDM, but close to unity for the relaxed state.
Asunto(s)
Fibras Musculares Esqueléticas/química , Animales , Fibras Musculares Esqueléticas/fisiología , Rana temporaria , Sarcómeros , Factores de Tiempo , Difracción de Rayos XRESUMEN
Amphiphiles are molecules such as surfactants or lipids that have a polar head group (hydrophilic) attached to nonpolar hydrophobic alkyl chains. Because of this characteristic they self-assemble in water and give rise to a wide range of phases with different structures and properties. Aqueous dispersions of amphiphiles are present in every aspect of day-to-day life-e.g., forming biological cell membranes, stabilizing emulsified food, or being used as soap. Time-resolved x-ray diffraction has been used to study the hexadecylhexa(oxyethylene glycol) ether (C16EO6)/water system, which shows an intermediate phase whose structure depends on the thermal path between lamellar and hexagonal structures. Heating the hexagonal phase from room temperature leads to a lamellar phase via an Ia3d cubic structure. Cooling from the lamellar phase initially leads epitaxially to an intermediate Rm before the hexagonal phase is reached. Both cubic and Rm phases are formed by very similar rod units, but the overall structures differ because of their spatial distribution and they both bridge morphologically the hexagonal and lamellar phases. The Ia3d does so on heating, whereas the Rm does on cooling. The structural path during the phase transitions is determined by topological similarities between the forming phase and the one from which it originates. Although the estimated curvature energies for these two phases are similar, on cooling, kinetics and topology are initial factors determining the path for the phase transitions, whereas on heating energy is the dominant factor.
Asunto(s)
Polietilenglicoles/química , Tensoactivos/química , Agua/química , Membrana Celular/fisiología , Modelos Moleculares , Difracción de Rayos X/métodosRESUMEN
The temperature dependence of the pressure-induced equilibrium unfolding of staphylococcal nuclease (Snase) was determined by fluorescence of the single tryptophan residue, FTIR absorption for the amide I' and tyrosine O-H bands, and small-angle X-ray scattering (SAXS). The results from these three techniques were similar, although the stability as measured by fluorescence was slightly lower than that measured by FTIR and SAXS. The resulting phase diagram exhibits the well-known curvature for heat and cold denaturation of proteins, due to the large decrease in heat capacity upon folding. The volume change for unfolding became less negative with increasing temperatures, consistent with a larger thermal expansivity for the unfolded state than for the folded state. Fluorescence-detected pressure-jump kinetics measurements revealed that the curvature in the phase diagram is due primarily to the rate constant for folding, indicating a loss in heat capacity for the transition state relative to the unfolded state. The similar temperature dependence of the equilibrium and activation volume changes for folding indicates that the thermal expansivities of the folded and transition states are similar. This, along with the fact that the activation volume for folding is positive over the temperature range examined, the nonlinear dependence of the folding rate constant upon temperature implicates significant dehydration in the rate-limiting step for folding of Snase.
Asunto(s)
Nucleasa Microcócica/química , Temperatura , Cinética , Cómputos Matemáticos , Presión , Pliegue de Proteína , Dispersión de Radiación , Espectrometría de Fluorescencia , Espectroscopía Infrarroja por Transformada de Fourier , Termodinámica , Rayos XRESUMEN
By using time-resolved X-ray diffraction, differential scanning calorimetry and scanning densitometry, we observed rapid formation at low temperature of a metastable ordered phase, termed LR1 phase, in fully hydrated dihexadecylphosphatidylethanolamine (DHPE). The LR1 phase has the same lamellar repeat period as the gel Lbeta phase but differs from the latter in its more ordered, orthorhombic hydrocarbon chain arrangement. It forms at about 12 degrees C upon cooling and manifests itself as splitting of the sharp, symmetric wide-angle X-ray peak of the DHPE gel phase into two reflections. This transition, designated the 'Y-transition', is readily reversible and proceeds with almost no hysteresis between cooling and heating scans. Calorimetrically, the LR1-->Lbeta transition is recorded as a low-enthalpy (0.2 kcal/mol) endothermic event. The formation of the LR1 phase from the gel phase is associated with a small, about 2 microl/g, decrease of the lipid partial specific volume recorded by scanning densitometry, in agreement with a volume calculation based on the X-ray data. The formation of the equilibrium Lc phase was found to take place from within the LR1 phase. This appears to be the only observable pathway for crystallisation of DHPE upon low-temperature incubation. Once formed, the Lc phase of this lipid converts directly into Lbeta phase at 50 degrees C, skipping the LR1 phase. Thus, the LR1 phase of DHPE can only be entered by cooling of the gel Lbeta phase. The data disclose certain similarities between the low-temperature polymorphism of DHPE and that of long-chain normal alkanes.
Asunto(s)
Membrana Dobles de Lípidos/química , Fosfatidiletanolaminas/química , Temperatura , Rastreo Diferencial de Calorimetría , Frío , Cristalización , Densitometría/métodos , Geles , Termodinámica , Difracción de Rayos X/métodosRESUMEN
The speed of the myofilament lattice spacing response to rapid changes in load or length of single, intact muscle fibres of the frog, was investigated during isometric tetani. During ramp releases at close to Vmax and during step length changes (completed within 250 microseconds), lattice spacing was calculated from the equatorial X-ray diffraction pattern (sampled at 250 microseconds time resolution using synchrotron radiation). Ramp releases (total shortening=1.39 %) caused a spacing increase, described with an exponential function (alpha=271 s-1, amplitude=1.15 nm) plus an elastic component having the time course of discharge of axial tension (amplitude 0.28 nm). For a step release (amplitude=0.87%), lattice expansion could be described with an exponential (alpha =1005 s-1, amplitude=0.56 nm) plus an elastic component of 0.25 nm amplitude. Lattice compression was associated with a step stretch (amplitude=0.62 %), and was also quasi-exponential (alpha=367 s-1, amplitude=0.74 nm), with an elastic component of 0.28 nm. The spacing change time course for length steps resembled that of the accompanying quick recovery of axial tension and the associated change in the meridional 14.5 nm reflection intensity, which are both believed to be determined by the kinetics of the molecular power stroke. Therefore, this shows that lattice spacing changes, arising from radial forces exerted by attached crossbridges, are fast enough to occur during the power stroke event.
Asunto(s)
Conformación Proteica , Animales , Miosinas/química , Rana temporaria , Factores de TiempoRESUMEN
The lamellar-to-hexagonal phase transition of a phospholipid/ surfactant mixed system of 1-palmitoyl-2-oleoyl-sn-glycero-3phosphocholine (POPC) and oligo(ethylene oxide)dodecyl ethers of type C12H25O(CH2CH2O)2H(C12E2) in molar surfactant/phospholipid ratio (RS/L) of 2 at low hydration driven by temperature has been studied by X-ray diffraction. The Lbeta-HII phase transition is a reversible two-state process showing hysteresis at fast temperature scan rates. The obtained hexagonal phase exhibits a temperature dependent structural change. The numbers of bound water molecules per composite particle (WS+L) absorbed in the lamellar and hexagonal phases are nearly the same, changing from WS+L = 5.0 to 4.7 during the phase transition. The fluidity of the alkyl chains on increasing the temperature and the close packing of the hydrophilic molecular parts are the driving parameters of the lamellar-to-hexagonal transformation. Copyright 1998 Academic Press.