Cell-Sized Liposomes and Droplets: Real-World Modeling of Living Cells

Hamada, Tsutomu; Yoshikawa, Kenichi

doi:10.3390/ma5112292

Cited by 55 publications

(42 citation statements)

References 54 publications

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“…consisting of mixtures of lipids and cholesterol have been used as model biomembranes [5][6][7] . In particular, studies of phase separation and membrane dynamics have been performed on such GUV consisting of saturated lipids, unsaturated lipids and cholesterol 8 .…”

Section: Introductionmentioning

confidence: 99%

Charge-induced phase separation in lipid membranes

et al. 2014

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The phase separation in lipid bilayers that include negatively charged lipids is examined experimentally. We observed phase-separated structures and determined the membrane miscibility temperatures in several binary and ternary lipid mixtures of unsaturated neutral lipid, dioleoylphosphatidylcholine (DOPC), saturated neutral lipid, dipalmitoylphosphatidylcholine (DPPC), unsaturated charged lipid, dioleoylphosphatidylglycerol (DOPG (-) ), saturated charged lipid, dipalmitoylphosphatidylglycerol (DPPG (-) ), and cholesterol. In binary mixtures of saturated and unsaturated charged lipids, the combination of the charged head with the saturation of hydrocarbon tail is a dominant factor for the stability of membrane phase separation. DPPG (-) enhances phase separation, while DOPG (-) suppresses it. Furthermore, the addition of DPPG (-) to a binary mixture of DPPC/cholesterol induces phase separation between DPPG (-) -rich and cholesterol-rich phases. This indicates that cholesterol localization depends strongly on the electric charge on the hydrophilic head group rather than on the ordering of the hydrocarbon tails. Finally, when DPPG (-) was added to a neutral ternary system of DOPC/DPPC/Cholesterol (a conventional model of membrane rafts), a three-phase coexistence was produced. We conclude by discussing some qualitative features of the phase behaviour in charged membranes using a free energy approach.

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Section: Introductionmentioning

confidence: 99%

Charge-induced phase separation in lipid membranes

et al. 2014

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“…This method is extremely simple and reproducible. Recently, the water-in-oil microdroplets are extended to form liposomes by using droplet-transfer method invented by Yoshikawa [51]. By using this approach, it is possible to modulate the lipid compositions of outer and inner leaflets and furthermore to orient a reconstituted membrane protein in liposomes [52].…”

Section: Reconstitution Of Membrane Protein Into Model Membranesmentioning

confidence: 99%

Reconstitution of Membrane Proteins into Model Membranes: Seeking Better Ways to Retain Protein Activities

Shen

Lithgow

Martin

2013

IJMS

108

104

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The function of any given biological membrane is determined largely by the specific set of integral membrane proteins embedded in it, and the peripheral membrane proteins attached to the membrane surface. The activity of these proteins, in turn, can be modulated by the phospholipid composition of the membrane. The reconstitution of membrane proteins into a model membrane allows investigation of individual features and activities of a given cell membrane component. However, the activity of membrane proteins is often difficult to sustain following reconstitution, since the composition of the model phospholipid bilayer differs from that of the native cell membrane. This review will discuss the reconstitution of membrane protein activities in four different types of model membrane—monolayers, supported lipid bilayers, liposomes and nanodiscs, comparing their advantages in membrane protein reconstitution. Variation in the surrounding model environments for these four different types of membrane layer can affect the three-dimensional structure of reconstituted proteins and may possibly lead to loss of the proteins activity. We also discuss examples where the same membrane proteins have been successfully reconstituted into two or more model membrane systems with comparison of the observed activity in each system. Understanding of the behavioral changes for proteins in model membrane systems after membrane reconstitution is often a prerequisite to protein research. It is essential to find better solutions for retaining membrane protein activities for measurement and characterization in vitro.

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“…This is the “semi-synthetic” approach (Luisi et al, 2006b; Stano et al, 2011), consisting in the construction of cell-like structures by encapsulating the minimal number of available DNA, RNA, proteins inside liposomes ( Figure 2 ). Currently this approach stems from the convergence of liposome technology and cell-free technology and it is investigated by several groups, as witnessed by the richness of the recently published original work (Budin et al, 2012; Hamada and Yoshikawa, 2012; Maeda et al, 2012; Kobori et al, 2013; Lentini et al, 2013; Nourian and Danelon, 2013). Several important concepts and technical issues play essential roles in semi-synthetic minimal cells, such as (1) life as an emergent property, (2) the minimal genome, (3) the production of functional water-soluble and membrane proteins, (4) the conditions for core-and-shell self-reproduction, (5) “spontaneous” versus “directed” preparation methods, and many others.…”

Section: Semi-synthetic Minimal Cellsmentioning

confidence: 99%

Chemical synthetic biology: a mini-review

Chiarabelli¹,

Stano²,

Luisi³

2013

Front. Microbiol.

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Chemical synthetic biology (CSB) is a branch of synthetic biology (SB) oriented toward the synthesis of chemical structures alternative to those present in nature. Whereas SB combines biology and engineering with the aim of synthesizing biological structures or life forms that do not exist in nature – often based on genome manipulation, CSB uses and assembles biological parts, synthetic or not, to create new and alternative structures. A short epistemological note will introduce the theoretical concepts related to these fields, whereas the text will be largely devoted to introduce and comment two main projects of CSB, carried out in our laboratory in the recent years. The “Never Born Biopolymers” project deals with the construction and the screening of RNA and peptide sequences that are not present in nature, whereas the “Minimal Cell” project focuses on the construction of semi-synthetic compartments (usually liposomes) containing the minimal and sufficient number of components to perform the basic function of a biological cell. These two topics are extremely important for both the general understanding of biology in terms of function, organization, and development, and for applied biotechnology.

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Cell-Sized Liposomes and Droplets: Real-World Modeling of Living Cells

Cited by 55 publications

References 54 publications

Charge-induced phase separation in lipid membranes

Charge-induced phase separation in lipid membranes

Reconstitution of Membrane Proteins into Model Membranes: Seeking Better Ways to Retain Protein Activities

Chemical synthetic biology: a mini-review

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