No abstract

No abstract

No abstract

We used x-ray and neutron diffraction to study the temperature- and pressure-dependent structure and phase behavior of the monoacylglycerides 1-monoelaidin (ME) and 1-monoolein (MO) in excess water. The monoacylglycerides were chosen for investigation of their phase behavior because they exhibit mesomorphic phases with one-, two-, and three-dimensional periodicity, such as lamellar, an inverted hexagonal and bicontinuous cubic phases, in a rather easily accessible temperature and pressure range. We studied the structure, stability, and transformations of the different phases over a wide temperature and pressure range, explored the epitaxial relations that exist between different phases, and established a relationship between the chemical structure of the lipid molecules and their phase behavior. For both systems, a temperature-pressure phase diagram has been determined in the temperature range from 0 to 100 degrees C at pressures from ambient up to 1400 bar, and drastic differences in phase behavior are found for the two systems. In MO-water dispersions, the cubic phase Pn3m extends over a large phase field in the T,p-plane. At temperatures above 95 degrees C, the inverted hexagonal phase is found. In the lower temperature region, a crystalline lamellar phase is induced at higher pressures. The phases found in ME-water include the lamellar crystalline Lc phase, the L beta gel phase, the L alpha liquid-crystalline phase, and two cubic phases belonging to the crystallographic space groups Im3m and Pn3m. In addition, the existence of metastable phases has been exploited. Between coexisting metastable cubic structures, a metric relationship has been found that is predicted theoretically on the basis of the curvature elastic energy approximation only.

The time course of structural changes accompanying the transition from the M412 intermediate to the BR568 ground state in the photocycle of bacteriorhodopsin (BR) from Halobacterium halobium was studied at room temperature with a time resolution of 15 ms using synchrotron radiation X‐ray diffraction. The M412 decay rate was slowed down by employing mutated BR Asp96Asn in purple membranes at two different pH‐values. The observed light‐induced intensity changes of in‐plane X‐ray reflections were fully reversible. For the mutated BR at neutral pH the kinetics of the structural alterations (tau 1/2 = 125 ms) were very similar to those of the optical changes characterizing the M412 decay, whereas at pH 9.6 the structural relaxation (tau 1/2 = 3 s) slightly lagged behind the absorbance changes at 410 nm. The overall X‐ray intensity change between the M412 intermediate and the ground state was about 9% for the different samples investigated and is associated with electron density changes close to helix G, B and E. Similar changes (tau 1/2 = 1.3–3.6 s), which also confirm earlier neutron scattering results on the BR568 and M412 intermediates trapped at ‐180 degrees C, were observed with wild type BR retarded by 2 M guanidine hydrochloride (pH 9.4). The results unequivocally prove that the tertiary structure of BR changes during the photocycle.

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.

The thermotropic phase behavior of zwitterionic/cationic binary lipid mixtures is investigated and compared to its corresponding lipidic phase diagram of mixtures complexed with DNA. We focus on isoelectric cationic lipid−DNA condensates where the number of cationic lipids equals the number of phosphate groups on the DNA. Using differential scanning calorimetry, X-ray scattering, freeze fracture electron microscopy, and film balance, we studied mixtures of di-myristoyl-phosphatidyl-choline (DMPC) and the cationic lipid, di-myristoyl-tri-methyl-ammonium-propane (DMTAP). The lipid phase diagram shows the well-known L α, L β ‘, and P β ‘ ripple phase with peritectic behavior at a low molar fraction of cationic lipid, χTAP < 0.12. Beyond χTAP = 0.8 crystalline phases appear. A systematic variation in the hydrocarbon chain tilt in the prevailing L β ‘ phase is measured by wide-angle X-ray scattering. Most importantly, the L β ‘ phase shows strong nonideal mixing with an azeotropic point at about 1:1 molar stoichiometry. This finding is related to the reduced headgroup area for equimolar mixtures found in monolayer pressure−area isotherms. The intercalation of DNA in cationic lipid−DNA complexes affects the lipid-phase behavior 2-fold:  (i) the chain-melting transition temperature shifts to higher temperatures and (ii) a demixing gap with coexistence of lipid vesicles and lipid−DNA complexes arises at a low cationic fraction, χTAP < 0.25. In agreement with experiments we present a thermodynamic model that describes the shift of the melting transition temperatures by DNA-induced electrostatic screening of the cationic membrane.

Lanyi (1990Lanyi ( , 1991a postulated an irreversible transition the FTIR spectra occur between M 2 and M G . This between M 1 and M 2 . This could not, however, hitherto suggests, that the tertiary structural changes between be unambiguously proven experimentally, although the M 1 and M 2 are responsible for the switch opening the presence of two M intermediates (M 1 and M 2 ) had been cytoplasmic half-channel of BR for reprotonation to deduced much earlier from spectroscopic measurements complete the catalytic cycle. These tertiary structural (Korenstein et al., 1978). changes seem to be triggered by a charge redistribution UV-VIS (Varo and Lanyi, 1991b;Varo et al., 1992; which might be a common feature of retinal proteins Zimanyi et al., 1992) and Fourier transform infrared also in signal transduction.(FTIR) spectroscopy (Ormos, 1991;Perkins et al., 1992) Keywords: conformational changes/hydration/ revealed a transition from M 1 to M 2 by a slight shift in M intermediates/photocycle/proton pumping the absorption maximum of the M intermediate and changes in the amide-I region (1650-1670 cm -1 ), respectively. Later, the FTIR results were, however, re-interpreted as giving no evidence for a M 1 to M 2 transition, due to