Constitutive plasma membrane targeting and microdomain localization of Dok5 studied by single-molecule microscopy

In the present study, single-molecule fluorescence microscopy was used to examine the characteristics of plasma membrane targeting and microdomain localization of enhanced yellow fluorescent protein (eYFP)-tagged wild-type Dok5 and its variants in living Chinese hamster ovary (CHO) cells. We found that Dok5 can target constitutively to the plasma membrane, and the PH domain is essential for this process. Furthermore, single-molecule trajectories analysis revealed that Dok5 can constitutively partition into microdomain on the plasma membrane. Finally, the potential mechanism of microdomain localization of Dok5 was discussed. This study provided insights into the characteristics of plasma membrane targeting and microdomain localization of Dok5 in living CHO cells. (C) 2008 Elsevier B.V. All rights reserved.

Interaction between alpha-actinin and negatively charged lipids membrane investigated by surface plasmon resonance and electrochemical methods

alpha-Actinin has been shown to be capable of interacting with some special membrane phospholipids directly, which is important for its function. In this study, hybrid bilayer membranes composed of negatively charged lipids are constructed on the surface plasmon resonance gold substrate and on the gold electrode, respectively, and the interaction between alpha-actinin and negatively charged lipids membrane is investigated by surface plasmon resonance, cyclic voltammetry and electrochemical impedance spectroscopy methods. alpha-Actinin is proved to be able to interact with the negatively charged lipids membrane directly. It can also insert at least partly into the membrane or lead to some defect or lesion in the membrane, which increase the permeability of the membrane. This study would bring some insight on the interaction between the alpha-actinin and the cell membranes in vivo.

The interaction of aldolase with the cytoplasmic domain of human erythrocyte band 3 inhibited by lanthanum ions

The effect of lanthanum ions on the activity of the cytoplasmic domain of human erythrocyte band 3 (CDB3), which was measured according to the inhibition to aldolase, was studied. In the presence of low concentration of lanthanum ions, the function of CDB3 to inhibit aldolase activity decreased significantly. It indicated that lanthanum ions in the erythrocyte would change the conformation of CDB3 and influence the control on aldolase activity.

SmC2P1, a C2 domain protein from Scophthalmus maximus that binds bacterial pathogen-infected lymphocytes and reduces bacterial survival

C2 domains are protein structural modules found in many eukaryotic proteins involved in signal transduction, membrane trafficking, and immune defense. Most of the studied C2 domain-containing proteins are multi-domained in structure, in which the C2 domain is an independently folded motif and plays an essential role in calcium-dependent membrane-targeting. Although C2 domains isolated from intact proteins have been studied for biological functions, no study on natural proteins containing C2 domain only has been documented. In this study, we identified a Scophthalmus maximus protein SmC2P1 that is comprised of a single C2 domain and lacks any other apparent domain structures. The deduced amino acid sequence of SmC2P1 contains 129 residues and shares 36-38% identities with the C2 domains of the perforins of several fish species. Like typical C2 domains, SmC2P1 is predicted to organize into eight beta-strands with a Ca2+-binding site located in inter-strand loops. SmC2P1 expression was detected, in deceasing order, in liver, spleen, blood, brain, muscle, kidney, gill, and heart. Experimental challenge of turbot with a bacterial pathogen significantly upregulated SmC2P1 expression in kidney in a time-dependent manner. Recombinant SmC2P1 purified from yeast exhibits no hemolytic activity but binds to pathogen-infected kidney lymphocytes in the presence of calcium. Furthermore, interaction of recombinant SmC2P1 with bacterium-infected lymphocytes reduced bacterial survival. These results indicate that SmC2P1 is a functional protein that is involved in host immune defense against bacterial infection. (C) 2010 Elsevier Ltd. All rights reserved.

Magnetic properties and magnetic domain structure of bulk glass forming Nd60Al10Fe20Co10 alloy

The transition from hard to soft magnetic behaviour with increasing quenching rate is shown for Nd60WAl10Fe20Co10 melt-spun ribbons with different thickness. Microstructure and magnetic domain structure of ribbons were studied by magnetic force microscopy (MFM). Particle sizes < 5 nm decreasing gradually with increasing quenching rate were deduced from topographic images which differ from large-scale magnetic domains with a periodicity of about 350 nm in all ribbons irrespective the coercivity. This indicates that the magnetic properties of the alloy are governed by interaction of small magnetic particles. It is concluded that the presence of short-range-ordered structures with a local ordering similar to the Al metastable Nd-Fe binary phase is responsible for the hard magnetic properties in samples subjected to relatively low quenching rate.

Domain structure of hard magnetic NdAlFeCo bulk metallic glass

Magnetic domain structure of hard magnetic Nd60Al10Fe20Co10 bulk metallic glass (BMG) has been studied by using magnetic force microscopy. In the magnetic force images it is shown that the exchange interaction type magnetic domains with a period of about 360 nm do exist in the BMG, which is believed to be associated with the appearance of hard-magnetic properties in this system. As the scale of the magnetic domain is much larger than the size of the short-range ordered atomic clusters existing in the BMG, it is believed that the large areas of magnetic contrast are actually a collection of a group of clusters aligned in parallel by strong exchange coupling interaction. After fully crystallization, the BMG exhibits paramagnetism. No obvious magnetic contrast is observed in the magnetic force images of fully crystallized samples, except for a small quantity of ferromagnetic crystalline phase with low coercivity and an average size of 900 nm.

Effects of Surface Microtopology and Membrane Stiffness on Kinetics of Selectin/Ligand interactions by a Modified BFP

More and more evidences come out to support that the functionality of adhesion molecules are influenced by the surface microtopology of cell carrier or substrate. Adhesive molecules usually express on the microvilli of a cell, providing a well-defined spatial configuration to mediate the adhesions to the counterpart molecules on the apposed surface.

Domain structure of a Nd60Al10Fe20Co10 bulk metallic glass

Magnetic domain structure of Nd60Al10Fe20Co10 bulk metallic glass (BMG) has been studied by using magnetic-force microscopy. In the magnetic-force images it is shown that the exchange-interaction-type magnetic domains with a period of about 360 nm do exist in the BMG, which is believed to be associated with the appearance of hard-magnetic properties in this system. The existence of the large-scale domains demonstrates that the magnetic moments of a great deal of short-scale ordered atomic clusters in the BMG have been aligned by exchange coupling. Annealing at 715 K leads to partial crystallization of the BMG. However, the exchange coupling is stronger in the annealed sample, which is considered to arise from the increase of transition-metal concentration in the amorphous phase due to the precipitation of Nd crystalline phase.

A time domain computation method for dynamic behavior of mooring system

A quasi-steady time domain method is developed for the prediction of dynamic behavior of a mooring system under the environmental disturbances, such as regular or irregular waves, winds and currents. The mooring forces are obtained in a static sense at each instant. The dynamic feature of the mooring cables can be obtained by incorporating the extended 3-D lumped-mass method with the known ship motion history. Some nonlinear effects, such as the influence of the instantaneous change of the wetted hull surface on the hydrostatic restoring forces and Froude-Krylov forces, are included. The computational results show a satisfactory agreement with the experimental ones.

Molecular Dynamics Simulation Of Adhesion Forces In A Dipalmitoylphosphatidylcholine Membrane

Adhesion forces of Dipalmitoylphosphatidylcholine ( DPPC) membrane in the gel phase are investigated by molecular dynamics ( MD) simulation. In the simulations, individual DPPC molecules are pulled out of DPPC membranes with different rates and we get the maximum adhesion forces of DPPC membrane. We find that the maximum adhesion forces increase with pull rate, from about 400 to 700 pN when pull rates are from 0.001 to 0.03 nm/ps. We analyze the relationship between pull rate and adhesion forces of different origins using Brownian dynamics and notice that viscosity of solvent plays an important role in adhesion forces. Then we simulate the motion of a single DPPC molecule in solvent and it elucidates that the maximum drag force is almost linear with respect to the pull rate. We use Stokes' relation to describe the motion of a single DPPC molecule and deduce the effective length of a DPPC molecule. Conformational analyses indicate that the free energy variation of a DPPC molecule inside and outside of the DPPC membrane is an essential part of adhesion energy.

Role Of Vertical Component Of Surface Tension Of The Droplet On The Elastic Deformation Of Pdms Membrane

Poly(dimethylsiloxane) (PDMS) has been widely used in lab-on-a-chip and micro- total analysis systems (mu-TAS), thus wetting and electrowetting behaviors of PDMS are of great importance in these devices. PDMS is a kind of soft polymer material, so the elastic deformation of PDMS membrane by a droplet cannot be neglected due to the vertical component of the interfacial tension between the liquid and vapor, and this vertical component of liquid-vapor surface tension is also balanced by the stress distribution within the PDMS membrane. Such elastic deformation and stress distribution not only affect the exact measurement of contact angle, but also have influence on the micro-fluidic behavior of the devices. Using ANSYS code, we simulated numerically the elastic deformation and stress distribution of PDMS membrane on a rigid substrate due to the liquid-vapor surface tension. It is found that the vertical elastic deformation of the PDMS membrane is on the order of several tens of nanometers due to the application of a droplet with a diameter of 2.31 mm, which is no longer negligible for lab-on-a-chip and mu-TAS. The vertical elastic deformation increases with the thickness of the PDMS membrane, and there exists a saturated membrane thickness, regarded as a semi-infinite membrane thickness, and the vertical elastic deformation reaches a limiting value when the membrane thickness is equal to or thicker than such saturated thickness. (C) Koninklijke Brill NV, Leiden, 2008.

Influence Of Different Amount Of Au On The Wetting Behavior Of Pdms Membrane

Polydimethylsiloxane (PDMS) has been widely used as a base material for bio-MEMS/NEMS devices. It is difficult for PDMS to transfer and spread aqueous solution as a kind of highly hydrophobic material. Therefore, surface modification is necessary for PDMS to make it hydrophilic. In this paper, a method of hydrophilization of PDMS surface is proposed. Gold is sputtered to the PDMS substrate by sputter coater in different average thicknesses. Relationship between the average thickness of gold on the PDMS substrate and the contact angle of the surface was studied. It was found that even gold of average thickness less than 1 nm can result in about 25 degrees change of contact angle. AFM is also used to get topographic information of PDMS surface coated with gold. Three cases are classified with different amount of Au: (1) Heterogeneous zone; (2) Transition zone; (3) Film zone. For heterogeneous zone, a simple model about heterogeneous phase wetting is put forward to interpret this phenomenon.

Energy principle of ferroelectric ceramics and single domain mechanical model

Many physical experiments have shown that the domain switching in a ferroelectric material is a complicated evolution process of the domain wall with the variation of stress and electric field. According to this mechanism, the volume fraction of the domain switching is introduced in the constitutive law of ferroelectric ceramic and used to study the nonlinear constitutive behavior of ferroelectric body in this paper. The principle of stationary total energy is put forward in which the basic unknown quantities are the displacement u (i) , electric displacement D (i) and volume fraction rho (I) of the domain switching for the variant I. Mechanical field equation and a new domain switching criterion are obtained from the principle of stationary total energy. The domain switching criterion proposed in this paper is an expansion and development of the energy criterion. On the basis of the domain switching criterion, a set of linear algebraic equations for the volume fraction rho (I) of domain switching is obtained, in which the coefficients of the linear algebraic equations only contain the unknown strain and electric fields. Then a single domain mechanical model is proposed in this paper. The poled ferroelectric specimen is considered as a transversely isotropic single domain. By using the partial experimental results, the hardening relation between the driving force of domain switching and the volume fraction of domain switching can be calibrated. Then the electromechanical response can be calculated on the basis of the calibrated hardening relation. The results involve the electric butterfly shaped curves of axial strain versus axial electric field, the hysteresis loops of electric displacement versus electric filed and the evolution process of the domain switching in the ferroelectric specimens under uniaxial coupled stress and electric field loading. The present theoretic prediction agrees reasonably with the experimental results given by Lynch.

Wave propagation and the frequency domain Green's functions in viscoelastic Biot/squirt (BISQ) media

In this paper, we examine the characteristics of elastic wave propagation in viscoelastic porous media, which contain simultaneously both the Biot-flow and the squirt-flow mechanisms (BISQ). The frequency-domain Green's functions for viscoelastic BISQ media are then derived based on the classic potential function methods. Our numerical results show that S-waves are only affected by viscoelasticity, but not by squirt-flows. However, the phase velocity and attenuation of fast P-waves are seriously influenced by both viscoelasticity and squirt-flows; and there exist two peaks in the attenuation-frequency variations of fast P-waves. In the low-frequency range, the squirt-flow characteristic length, not viscoelasticity, affects the phase velocity of slow P-waves, whereas it is opposite in the high-frequency range. As to the contribution of potential functions of two types of compressional waves to the Green's function, the squirt-flow length has a small effect, and the effects of viscoelastic parameter are mainly in the higher frequency range. Crown Copyright (C) 2006 Published by Elsevier Ltd. All rights reserved.

Experimental study on the microstructure and nanomechanical properties of the wing membrane of dragonfly

Detailed investigations on the microstructure and the mechanical properties of the wing membrane of the dragonfly are carried out. It is found that in the direction of the thickness the membrane was divided into three layers rather than a single entity as traditionally considered, and on the surfaces the membrane displays a random distribution rough microstructure that is composed of numerous nanometer scale columns coated by the cuticle wax secreted. The characteristics of the surface structure are measured and described. The mechanical properties of the membranes taken separately from the wings of live and dead dragonflies are investigated by the nanoindentation technique. The Young's moduli obtained here are approximately two times greater than the previous result, and the reasons that yield the difference are discussed.