scholarly journals Calcium‐Mediated Liposome Fusion to Engineer Giant Lipid Vesicles with Cytosolic Proteins and Reconstituted Mammalian Proteins

2020 ◽  
Vol 4 (11) ◽  
pp. 2000153
Author(s):  
Yannick R. F. Schmid ◽  
Leo Scheller ◽  
Sebastian Buchmann ◽  
Petra S. Dittrich
Keyword(s):  
Lipid Vesicles ◽  
Cytosolic Proteins ◽  
Mammalian Proteins ◽  
Liposome Fusion

The structure of membranes and membrane proteins embedded in amorphous ice

1992 ◽  
Vol 50 (1) ◽  
pp. 432-433
Author(s):  
Uwe Lücken ◽  
Joachim Jäger
Keyword(s):  
Phase Transition ◽  
Transition Temperature ◽  
Membrane Proteins ◽  
Phase Transition Temperature ◽  
Lipid Vesicles ◽  
Freezing Stress ◽  
Amorphous Ice ◽  
Different Temperatures ◽  
Band Pass ◽  
Lipid Polymorphism

TEM imaging of frozen-hydrated lipid vesicles has been done by several groups Thermotrophic and lyotrophic polymorphism has been reported. By using image processing, computer simulation and tilt experiments, we tried to learn about the influence of freezing-stress and defocus artifacts on the lipid polymorphism and fine structure of the bilayer profile. We show integrated membrane proteins do modulate the bilayer structure and the morphology of the vesicles.Phase transitions of DMPC vesicles were visualized after freezing under equilibrium conditions at different temperatures in a controlled-environment vitrification system. Below the main phase transition temperature of 24°C (Fig. 1), vesicles show a facetted appearance due to the quasicrystalline areas. A gradual increase in temperature leads to melting processes with different morphology in the bilayer profile. Far above the phase transition temperature the bilayer profile is still present. In the band-pass-filtered images (Fig. 2) no significant change in the width of the bilayer profile is visible.

Anticoagulant Activity of β2-Glycoprotein I Is Potentiated by a Distinct Subgroup of Anticardiolipin Antibodies

1992 ◽  
Vol 68 (03) ◽  
pp. 297-300 ◽  
Author(s):  
Monica Galli ◽  
Paul Comfurius ◽  
Tiziano Barbui ◽  
Robert F A Zwaal ◽  
Edouard M Bevers
Keyword(s):  
Human Plasma ◽  
Anticoagulant Activity ◽  
Factor Xa ◽  
Type A ◽  
Lipid Vesicles ◽  
Dependent Manner ◽  
Type B ◽  
Distinct Subgroup ◽  
Glycoprotein I ◽  
Thrombin Formation

SummaryPlasmas of 16 patients positive for both IgG anticardiolipin (aCL) antibodies and lupus anticoagulant (LA) antibodies were subjected to adsorption with liposomes containing cardiolipin. In 5 of these plasmas both the anticardiolipin and the anticoagulant activities were co-sedimented with the liposomes in a dose-dependent manner, whereas in the remaining cases only the anticardiolipin activity could be removed by the liposomes, leaving the anticoagulant activity (LA) in the supernatant plasma. aCL antibodies purified from the first 5 plasmas were defined as aCL-type A, while the term aCL-type B was used for antibodies in the other 11 plasmas, from which 2 were selected for this study.Prolongation of the dRVVT was produced by affinity-purified aCL-type A antibodies in plasma of human as well as animal (bovine, rat and goat) origin. aCL-type B antibodies were found to be devoid of anticoagulant activity, while the corresponding supernatants containing LA IgG produced prolongation of the dRVVT only in human plasma.These anticoagulant activities of aCL-type A and of LA IgG's were subsequently evaluated in human plasma depleted of β2-glycoprotein I (β2-GPI), a protein which was previously shown to be essential in the binding of aCL antibodies to anionic phospholipids. Prolongation of the dRVVT by aCL-type A antibodies was abolished using β2-GPI deficient plasma, but could be restored upon addition of β2-GPI. In contrast, LA IgG caused prolongation of the dRVVT irrespective of the presence or absence of β2-GPI.Since β2-GPI binds to negatively-charged phospholipids and impedes the conversion of prothrombin by the factor Xa/Va enzyme complex (Nimpf et al., Biochim Biophys Acta 1986; 884: 142–9), comparison was made of the effect of aCL-type A and aCL-type B antibodies on the rate of thrombin formation in the presence and absence of β2-GPI. This was measured in a system containing highly purified coagulation factors Xa, Va and prothrombin and lipid vesicles composed of 20 mole% phosphatidylserine and 80 mole% phosphatidylcholine. No inhibition on the rate of thrombin formation was observed with both types of aCL antibodies when either β2-GPI or the lipid vesicles were omitted. Addition of β2-GPI to the prothrombinase assay in the presence of lipid vesicles causes a time-dependent inhibition which was not affected by the presence of aCL-type B or non-specific IgG. In contrast, the presence of aCL-type A antibodies dramatically increased the anticoagulant effect of β2-GPI. These data indicate that the anticoagulant activity of aCL-type A antibodies in plasma is mediated by β2-GPI.

Clathrin-Inspired Coating and Stabilization of Liposomes by DNA Self-Assembly

2019 ◽  
Author(s):  
Kevin N. Baumann ◽  
Luca Piantanida ◽  
Javier García-Nafría ◽  
Diana Sobota ◽  
Kislon Voïtchovsky ◽  
...  
Keyword(s):  
Self Assembly ◽  
Mechanical Stability ◽  
Lipid Vesicles ◽  
The Self ◽  
Force Microscopy ◽  
Atomic Force ◽  
Cryo Electron Microscopy ◽  
Further Development ◽  
Liposome Surface ◽  
Dna Coating

The self-assembly of the protein clathrin on biological membranes facilitates essential processes of endocytosis in biological systems and has provided a source of inspiration for materials design by the highly ordered structural appearance. By mimicking the architecture of clathrin self-assemblies to coat liposomes with biomaterials, new classes of hybrid carriers can be derived. Here we present a method for fabricating DNA-coated liposomes by hydrophobically anchoring and subsequently growing a DNA network on the liposome surface which structurally mimics clathrin assemblies. Dynamic light scattering (DLS), ζ-potential and cryo-electron microscopy (cryo-EM) measurements independently demonstrate successful DNA coating. Nanomechanical measurements conducted with atomic force microscopy (AFM) show that the DNA coating enhances the mechanical stability of the liposomes relative to uncoated ones. Furthermore, we provide the possibility to reverse the coating process by triggering the disassembly of the DNA coating through a toehold-mediated displacement reaction. Our results describe a straightforward, versatile, and reversible approach for coating and stabilizing lipid vesicles by an interlaced DNA network. This method has potential for further development towards the ordered arrangement of tailored functionalities on the surfaces of liposomes and for applications as hybrid nanocarrier.

Efficient Delivery of Cytosolic Proteins by Protein-Hexahistidine-Metal Co-Assemblies

2021 ◽  
Author(s):  
Wenjuan Huang ◽  
Sijie Zhou ◽  
Bojiao Tang ◽  
Hongyan Xu ◽  
Xiaoxiao Wu ◽  
...  
Keyword(s):  
Cytosolic Proteins ◽  
Efficient Delivery

Binding Assay of Cytosolic Proteins to the Cytoskeleton

BIO-PROTOCOL ◽  
2013 ◽  
Vol 3 (22) ◽  
Author(s):  
Stefano Del Duca ◽  
Giampiero Cai
Keyword(s):  
Binding Assay ◽  
Cytosolic Proteins

scholarly journals Discontinuous DNA synthesis by purified mammalian proteins.

1990 ◽  
Vol 265 (30) ◽  
pp. 18461-18471 ◽  
Author(s):  
M Goulian ◽  
S H Richards ◽  
C J Heard ◽  
B M Bigsby
Keyword(s):  
Dna Synthesis ◽  
Mammalian Proteins

scholarly journals Formation of Giant Lipid Vesicles in the Presence of Nonelectrolytes—Glucose, Sucrose, Sorbitol and Ethanol

Processes ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 945
Author(s):  
Qiong Wang ◽  
Ning Hu ◽  
Jincan Lei ◽  
Qiurong Qing ◽  
Jing Huang ◽  
...  
Keyword(s):  
Sodium Chloride ◽  
Cell Membrane ◽  
Lipid Membranes ◽  
Volume Fraction ◽  
Hydrodynamic Force ◽  
Lipid Vesicles ◽  
Ethanol Solution ◽  
Sugar Solution ◽  
Ethanol Content ◽  
Membrane Models

Lipid vesicles, especially giant lipid vesicles (GLVs), are usually adopted as cell membrane models and their preparation has been widely studied. However, the effects of some nonelectrolytes on GLV formation have not been specifically studied so far. In this paper, the effects of the nonelectrolytes, including sucrose, glucose, sorbitol and ethanol, and their coexistence with sodium chloride, on the lipid hydration and GLV formation were investigated. With the hydration method, it was found that the sucrose, glucose and sorbitol showed almost the same effect. Their presence in the medium enhanced the hydrodynamic force on the lipid membranes, promoting the GLV formation. GLV formation was also promoted by the presence of ethanol with ethanol volume fraction in the range of 0 to 20 percent, but higher ethanol content resulted in failure of GLV formation. However, the participation of sodium chloride in sugar solution and ethanol solution stabilized the lipid membranes, suppressing the GLV formation. In addition, the ethanol and the sodium chloride showed the completely opposite effects on lipid hydration. These results could provide some suggestions for the efficient preparation of GLVs.

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