Abstract
Background: The transformation of the supported lipid bilayer (SLB) membrane by extracted cytosol from living resources, has recently drawn much attention. It enables us to address the question of whether the purified phospholipid SLB membrane, including lipids related to amoeba locomotion, which was discussed in many previous studies, exhibits membrane deformation in the presence of cytosol extracted from amoeba; Methods: In this report, a method for reconstituting a supported lipid bilayer (SLB) membrane, composed of purified phospholipids and cytosol extracted from Dictyostelium discoideum, is described. This technique is a new reconstitution method combining the artificial constitution of membranes with the reconstitution using animate cytosol (without precise purification at a molecular level), contributing to membrane deformation analysis; Results: The morphology transition of a SLB membrane composed of phosphatidylcholines, after the addition of cytosolic extract, was traced using a confocal laser scanning fluorescence microscope. As a result, pore formation in the SLB membrane was observed and phosphatidylinositides incorporated into the SLB membrane tended to suppress pore formation and expansion; Conclusions: The current findings imply that phosphatidylinositides have the potential to control cytoplasm activity and bind to a phosphoinositide-containing SLB membrane.
Highlights
Reconstitution approaches for cell motility have drawn much attention over recent decades.Prof. van Oudenaarden’s group reported that lipid vesicles coated with the Listeria monocytogenes virulence protein ActA, are propelled by in vitro actin polymerization [1]
Given that the purified lipid supported lipid bilayer (SLB) membrane artificially constituted and the extracted cytosol[14], is reconstituted, is a novel reconstitution method.is Here, based on the Noireaux group’s work we develop this a new current technique is using a novelcytosol reconstitution method
Including lipids related to amoeba was discussed in many previous the purified phospholipid SLB membrane. including lipids related to amoeba locomotion, which was studies, exhibits membrane deformation in the presence of cytosol extracted from amoeba
Summary
Reconstitution approaches for cell motility have drawn much attention over recent decades. In another report, atomic force microscopy demonstrated that nanometer-sized pores were an SLB membrane containing phosphatidylinositol-4,5-bisphosphate can lead to the self-assembly of formed in the pure phospholipid SLB membrane, in the presence of the Escherichia coli cytosol [14]. 1, and the Arp2/3 complex, as well as uniform and short polymerized actin [15] These reports fromthat prokaryotic cells or purified proteins related to cell motion or dynamics, is a powerful tool this technique, which we call the novel reconstitution method, using a purified lipid SLB for measuring the cytosol energyfrom and prokaryotic kinetics of acells membrane transition. Dictyostelium discoideum, a new reconstitution method SLB using cytosol extracted from the eukaryotic amoeba and we by develop generating a purified phospholipid membrane on a glass substrate using the vesicle.
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