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P Krüger

Publications and source records attributed to P Krüger.

At least 19 recordsLinked to original sources
PubMed2001

Giant lateral electrostriction in ferroelectric liquid-crystalline elastomers.

Mechanisms for converting electrical energy into mechanical energy are essential for the design of nanoscale transducers, sensors, actuators, motors, pumps, artificial muscles, and medical microrobots. Nanometre-scale actuation has to date been mainly achieved by using the (linear) piezoelectric effect in certain classes of crystals (for example, quartz), and 'smart' ceramics such as lead zirconate titanate. But the strains achievable in these materials are small--less than 0.1 per cent--so several alternative materials and approaches have been considered. These include grafted polyglutamates (which have a performance comparable to quartz), silicone elastomers (passive material--the constriction results from the Coulomb attraction of the capacitor electrodes between which the material is sandwiched) and carbon nanotubes (which are slow). High and fast strains of up to 4 per cent within an electric field of 150 MV x m(-1) have been achieved by electrostriction (this means that the strain is proportional to the square of the applied electric field) in an electron-irradiated poly(vinylidene fluoride-trifluoroethylene) copolymer. Here we report a material that shows a further increase in electrostriction by two orders of magnitude: ultrathin (less than 100 nanometres) ferroelectric liquid-crystalline elastomer films that exhibit 4 per cent strain at only 1.5 MV x m(-1). This giant electrostriction was obtained by combining the properties of ferroelectric liquid crystals with those of a polymer network. We expect that these results, which can be completely understood on a molecular level, will open new perspectives for applications.

Journal Article↗
PubMed2001

Extending the capabilities of targeted molecular dynamics: simulation of a large conformational transition in plasminogen activator inhibitor 1.

Plasminogen activator inhibitor type 1 (PAI-1) is an inhibitor of plasminogen activators such as tissue-type plasminogen activator or urokinase-type plasminogen activator. For this molecule, different conformations are known. The inhibiting form that interacts with the proteinases is called the active form. The noninhibitory, noncleavable form is called the latent form. X-ray and modeling studies have revealed a large change in position of the reactive center loop (RCL), responsible for the interaction with the proteinases, that is inserted into a beta-sheet (s4A) in the latent form. The mechanism underlying this spontaneous conformational change (half-life = 2 h at 37 degrees C) is not known in detail. This investigation attempts to predict a transition path from the active to the latent structure at the atomic level, by using simulation techniques. Together with targeted molecular dynamics (TMD), a plausible assumption on a rigid body movement of the RCL was applied to define an initial guess for an intermediate. Different pathways were simulated, from the active to the intermediate, from the intermediate to the latent structure and vice versa under different conditions. Equilibrium simulations at different steps of the path also were performed. The results show that a continuous pathway from the active to the latent structure can be modeled. This study also shows that this approach may be applied in general to model large conformational changes in any kind of protein for which the initial and final three-dimensional structure is known.

Computer Simulation↗
PubMed2000

Submolecular organization of DMPA in surface monolayers: beyond the two-layer model.

A new approach to the data refinement of X-ray reflection measurements from lipid surface monolayers, applied to DMPA on pure water, reveals the structural organization of the lipid in unprecedented detail and provides new insights into headgroup conformation and hydration as a function of lateral pressure. While conventional box models are incapable of modeling the experimental data at high momentum transfer satisfactorily, a quasimolecular composition-space refinement approach using distribution functions to map the spatial organization of submolecular headgroup fragments yields a much better description and overcomes inherent difficulties of box models. Upon going from the fluid liquid-expanded (LE) phase to the hexatic liquid-condensed (LC) phase, the orientation of the headgroup is tightly coupled to the ordering of the acyl chains. Headgroups tilt toward the surface normal to accommodate for the large reduction in available area per lipid molecule. The spread of the headgroup fragment distribution is considerably larger than the global interface roughness and increases slightly with compression. In distinction to earlier work on DMPE using the two-box approach, we find that the phosphate hydration stays essentially constant across the whole isotherm. The discrepancy between the results observed with the different models is attributed to intrinsic deficiencies of the box model.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed2000

Value of myofibrillar protein catabolic rate in Duchenne muscular dystrophy. A study after lower limb surgery.

Previous studies have shown a decreased progression of the course in Duchenne muscular dystrophy (DMD) patients treated by lower limb surgery in early childhood. The use of 3-methylhistidine (3-MH) excretion and 3-MH/creatinine excretion ratio as an appropriate indicator for the myofibrillar protein catabolic rate (MPCR) in muscle disorders is discussed controversially. To explore this issue we studied the renal excretion of (3-MH) over a period up to 24 months (on average 20.8 +/- 1.4) after operation in 15 consecutive DMD patients with an average age at operation of 8.75 (+/- 2.43 years) to evaluate the myofibrillar protein catabolic rate. No significant change of the MPCR could be found in our population over the follow-up period. However, the formula for the calculation of the MPCR contains quantities which are not precisely known in DMD or assumed to be constant over the progressive course of DMD. Summarizing MPCR cannot be recommended for the assessment of therapeutic efficacy in DMD.

Adolescent↗
PubMed2000

Molecular chirality and domain shapes in lipid monolayers on aqueous surfaces.

The shapes of domain boundaries in the mesoscopic phase separation of phospholipids in aqueous surface monolayers are analyzed with particular attention to the influence of molecular chirality. We have calculated equilibrium shapes of such boundaries, and show that the concept of spontaneous curvature-derived from an effective pair potential between the chiral molecules-yields an adequate description of the contribution of chirality to the total energy of the system. For enantiomeric dipalmitoylphosphatidylcholine in pure monolayers, and in mixtures with impurities that adsorb preferentially at the (one-dimensional) boundary line between the isotropic and anisotropic fluid phases, such as cyanobiphenyl (5CB), a total energy term that includes line tension, electrostatic dipole-dipole interaction, and spontaneous curvature is sufficient to describe the shapes of well-separated domain boundaries in full detail. As soon as interdomain distances fall below the domain sizes upon compression of a monolayer, fluctuations take over in determining its detailed structural morphology. Using Minkowski measures for the well-studied dimyristoyl phosphatidic acid (DMPA)/cholesterol system, we show that calculations accounting for line tension, electrostatic repulsion, and molecular chirality yield boundary shapes that are of the same topology as the experimentally observed structures. At a fixed molecular area in the phase coexistence region, the DMPA/cholesterol system undergoes an exponential decay of the line tension lambda with decreasing subphase temperature T.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1999

Effect of hydrophobic surfactant peptides SP-B and SP-C on binary phospholipid monolayers. I. Fluorescence and dark-field microscopy.

The influence of the hydrophobic proteins SP-B and SP-C, isolated from pulmonary surfactant, on the morphology of binary monomolecular lipid films containing phosphocholine and phosphoglycerol (DPPC and DPPG) at the air-water interface has been studied using epifluorescence and dark-field microscopy. In contrast to previously published studies, the monolayer experiments used the entire hydrophobic surfactant protein fraction (containing both the SP-B and SP-C peptides) at physiologically relevant concentrations (approximately 1 wt %). Even at such low levels, the SP-B/C peptides induce the formation of a new phase in the surface monolayer that is of lower intrinsic order than the liquid condensed (LC) phase that forms in the pure lipid mixture. This presumably leads to a higher structural flexibility of the surface monolayer at high lateral pressure. Variation of the subphase pH indicates that electrostatic interaction dominates the association of the SP-B/C peptides with the lipid monolayer. As evidenced from dark-field microscopy, monolayer material is excluded from the DPPC/DPPG surface film on compression and forms three-dimensional, surface-associated structures of micron dimensions. Such exclusion bodies formed only with SP-B/C peptides. This observation provides the first direct optical evidence for the squeeze-out of pulmonary surfactant material in situ at the air-water interface upon increasing monolayer surface pressures.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1998

Analysis of protein self-association at constant concentration by fluorescence-energy transfer.

Fluorescence-resonance-energy transfer from subunits labelled with a fluorescence donor group to subunits labelled with a fluorescence acceptor group can be used for quantitative analysis of protein self-association. The present approach evaluates fluorescence measurements on mixtures of equimolar solutions of donor-labelled and acceptor-labelled protein composed by systematic variation of the volume ratio. Its attractive feature is that it allows the determination of equilibrium constants at fixed total concentration. Problems encountered by most other methods, which require the equilibria to be followed to high dilution, are avoided. Conditions to be fulfilled are that a reactive site is available on the protein for specific introduction of the labels and that labelling neither affects the conformation nor interferes with the intermolecular interactions. It is desirable that the Forster distance of the donor/acceptor pair complies with its separation. While dimerisation constants can be determined exclusively by fluorescence measurements, the analysis of more complex cases of self-association depends on additional independent information. This communication reports on an application of the approach to the association/dissociation equilibrium between insulin monomers and dimers. Labelling of insulin at the epsilon-amino group of LysB29 does not disturb the conformation nor does it affect dimerisation. 2-Aminobenzoyl and 3-nitrotyrosyl residues served as the donor/acceptor pairs. Because they are less bulky than most other fluorescence labels and are of balanced polarity they do not alter the chemical nature of the protein. Their Forster distance of 29 A matches their 32-A separation in the insulin dimer. Energy transfer was measured as a function of the molar fractions of donor-insulin and acceptor-insulin at constant total concentration. Evaluation of this dependence resulted in a dimerisation constant, K12, of 0.72x10(5) M(-1). Its agreement with values obtained with other methods demonstrates that the present approach is a reliable alternative.

Circular Dichroism↗
PubMed1997

Calculations of the CD spectrum of bovine pancreatic ribonuclease.

CD spectra of bovine pancreatic ribonuclease A (RNase A) and its subtilisin-modified from (RNase S) have been calculated, based upon high-resolution structures from x-ray diffraction. All known transitions in the peptide and side-chain groups, especially the aromatic and disulfide groups, have been included. Calculations have been performed with both the matrix method and with first-order perturbation theory. A newly developed method for treating the electrostatic interactions among transition charge densities and between static charge distributions and transition charge densities is used. The effects of local electrostatic fields upon the group transition energies are included for all transitions. Rotational strengths generated by the matrix method were combined with Gaussian band shapes to generate theoretical CD spectra. The calculated spectra reproduce the signs and approximate magnitudes of the near-uv CD bands of both RNase A and S. Agreement is most satisfactory for the negative 275 nm band, dominated by tyrosine contributions. In agreement with two previous studies by other workers, coupling between Tyr 73 and Tyr 115 is the single most important factor in this band. The positive band observed near 240 nm is dominated by disulfide contributions, according to our results. The far-uv CD spectrum is poorly reproduced by the calculations. The observed 208 nm band, characteristic of alpha-helices, is absent from the calculated spectrum, probably because the helices in RNase are short. A strong positive couplet centered near 190 nm is predicted but not observed. Possible reasons for these incorrect predictions of the current theoretical model in the far-uv are discussed.

Animals↗
PubMed1996

Simulation of a complex protein structural change: the T <--> R transition in the insulin hexamer.

The T <--> R transition in the insulin hexamer is an outstanding model for protein structural changes in terms of its extent and complexity: the limiting structures T(6), T(3)R(3) and R(6) have been defined by X-ray crystallography. The transition occurs cooperatively within trimers. It involves displacements of >30 A and a secondary structural rearrangement of 15% of the peptide chain between extended and helical conformations. Experimental data for the transition are plentiful. Theoretical methods to simulate pathways without constraints would never succeed with such substantial transitions. We have developed two approaches, targeted energy minimization (TEM) and targeted molecular dynamics (TMD). Previously successful in simulating the T <--> R transition of the insulin monomer, these procedures are also shown here to be effective in the hexamer. With TMD, more conformational space is explored and pathways are found at 500 kJ/mol lower energy than with TEM. Because the atoms have to meet distance constraints in sum rather than individually, a high degree of conformational freedom and independence is implied. T(6) --> T(3)R(3) and T(3)R(3) --> T(6) pathways do not coincide because the transformation is directed. One subunit enters a dead end pathway in one direction of the TMD simulation, which shows that constraint and freedom are critically balanced. The ensemble of productive pathways represents a plausible corridor for the transition. A video display of the transformations is available.

Crystallography, X-Ray↗