Manipulation and sorting of membrane proteins using patterned diffusion-aided ratchets with AC fields in supported lipid bilayers

Cheetham, Matthew R.; Bramble, Jonathan P.; McMillan, Duncan G. G.; Bushby, Richard J.; Olmsted, Peter D.; Jeuken, Lars J. C.; Evans, Stephen D.

doi:10.1039/c2sm25473e

Cited by 21 publications

(18 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Recently, we have demonstrated that charged lipids and a “simple” transmembrane protein can be concentrated in SLBs by using a “fish-trap” structure in conjunction with an ac electric field . We have also shown the directed transport and orientational sorting of transmembrane proteins by using double-sawtooth ratchet patterns . However, our previous work has two major shortcomings.…”

mentioning

confidence: 96%

On-Chip Alternating Current Electrophoresis in Supported Lipid Bilayer Membranes

et al. 2012

Self Cite

View full text Add to dashboard Cite

By forming lipid bilayers within SU8 patterns, between interdigitated electrodes, we have demonstrated that it is possible to manipulate charged membrane components using low applied voltages over relatively short time scales. Two distinct patterns were studied: a nested "fish trap" which served as a molecular trap, and a diffusion aided Brownian ratchet which operated as a molecular pump. By reducing the size of the structures we have demonstrated that large applied fields (>200 V/cm) can be achieved on-chip, using low applied potentials (<13 V). By using ac fields applied orthogonal to the direction of desired motion, the molecular pumps provide a voltage independent method for moving charged components within lipid membranes over large distances. The reduced scale of the trap structures compared to those previously used in our laboratory has led to over a 10-fold decrease in the operational time require for charge build-up, from 16 h down to 1.5 h. The observed benefits of scaling means that these systems should be suitable for the on-chip separation and manipulation of charged species within supported lipid membranes.

show abstract

mentioning

confidence: 96%

On-Chip Alternating Current Electrophoresis in Supported Lipid Bilayer Membranes

et al. 2012

Self Cite

View full text Add to dashboard Cite

show abstract

“…The polymeric bilayer scaffolds that confine the hybrid thylakoid membrane can potentially enhance the membrane stability by supporting the fluid membrane from the side. Furthermore, the framework would be utilized to transport and concentrate the membrane-bound components by applying electric fields. , In the present study, we incorporated the whole thylakoid membrane, but it would be interesting to incorporate appressed (grana) and nonappressed (stroma lamellae) thylakoid membranes separately for concentrating some of the membrane components. At the same time, there are some technical limitations present in the patterned thylakoid membrane.…”

Section: Discussionmentioning

confidence: 99%

Photosynthetic Model Membranes of Natural Plant Thylakoid Embedded in a Patterned Polymeric Lipid Bilayer

et al. 2020

View full text Add to dashboard Cite

Thylakoid membranes in the chloroplast of plants, algae, and cyanobacteria are the powerhouse of photosynthesis, capturing solar energy and converting it into chemical energy. Although their structures and functions have been extensively studied, the intrinsically heterogeneous and dynamic nature of the membrane structures is still not fully understood. Investigating native thylakoid membranes in vivo is difficult due to their small size and limited external access to the chloroplast interior, while the bottom-up approaches based on model systems have been hampered by the sheer complexity of the native membrane. Here, we try to fill the gap by reconstituting the whole thylakoid membrane into a patterned substrate-supported planer bilayer. A mixture of thylakoid membrane purified from spinach leaves and synthetic phospholipid 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) vesicles spontaneously formed a laterally continuous and fluid two-dimensional (2D) membrane in the scaffold of the patterned polymeric bilayer. Chlorophyll fluorescence arising from photosystem II (PSII) recovered after photobleaching, suggesting that the membrane components are laterally mobile. The reversible changes of chlorophyll fluorescence in the presence of the electron acceptors and/or inhibitors indicated that the electron transfer activity of PSII was retained. Furthermore, we confirmed the electron transfer activity of photosystem I (PSI) by observing the generation of nicotinamide adenine dinucleotide phosphate (NADPH) in the presence of water-soluble ferredoxin and ferredoxin−NADP + reductase. The lateral mobility of membrane-bound molecules and the functional reconstitution of major photosystems provide evidence that our hybrid thylakoid membranes could be an excellent experimental platform to study the 2D molecular organization and machinery of photosynthesis.

show abstract

“…Yoshina-Ishii and Boxer continued this work by showing that it was possible to manipulate lipids within membrane arrays . More recently, Cheetham and Roth and co-workers published a series of papers in which ratchet structures were fabricated by photolithography and microcontact printing for the movement and concentration of both lipids and membrane proteins within SLBs. − …”

Section: Introductionmentioning

confidence: 99%

Simple, Direct Routes to Polymer Brush Traps and Nanostructures for Studies of Diffusional Transport in Supported Lipid Bilayers

et al. 2017

Self Cite

View full text Add to dashboard Cite

Patterned poly(oligo ethylene glycol) methyl ether methacrylate (POEGMEMA) brush structures may be formed by using a combination of atom-transfer radical polymerization (ATRP) and UV photopatterning. UV photolysis is used to selectively dechlorinate films of 4-(chloromethyl)phenyltrichlorosilane (CMPTS) adsorbed on silica surfaces, by exposure either through a mask or using a two-beam interferometer. Exposure through a mask yields patterns of carboxylic acid-terminated adsorbates. POEGMEMA may be grown from intact Cl initiators that were masked during exposure. Corrals, traps, and other structures formed in this way enable the patterning of proteins, vesicles, and, following vesicle rupture, supported lipid bilayers (SLBs). Bilayers adsorbed on the carboxylic acid-terminated surfaces formed by C–Cl bond photolysis in CMPTS exhibit high mobility. SLBs do not form on POEGMEMA. Using traps consisting of carboxylic acid-functionalized regions enclosed by POEGMEMA structures, electrophoresis may be observed in lipid bilayers containing a small amount of a fluorescent dye. Segregation of dye at one end of the traps was measured by fluorescence microscopy. The increase in the fluorescence intensity was found to be proportional to the trap length, while the time taken to reach the maximum value was inversely proportional to the trap length, indicating uniform, rapid diffusion in all of the traps. Nanostructured materials were formed using interferometric lithography. Channels were defined by exposure of CMPTS films to maxima in the interferogram, and POEGMEMA walls were formed by ATRP. As for the micrometer-scale patterns, bilayers did not form on the POEGMEMA structures, and high lipid mobilities were measured in the polymer-free regions of the channels.

show abstract

Manipulation and sorting of membrane proteins using patterned diffusion-aided ratchets with AC fields in supported lipid bilayers

Cited by 21 publications

References 29 publications

On-Chip Alternating Current Electrophoresis in Supported Lipid Bilayer Membranes

On-Chip Alternating Current Electrophoresis in Supported Lipid Bilayer Membranes

Photosynthetic Model Membranes of Natural Plant Thylakoid Embedded in a Patterned Polymeric Lipid Bilayer

Simple, Direct Routes to Polymer Brush Traps and Nanostructures for Studies of Diffusional Transport in Supported Lipid Bilayers

Contact Info

Product

Resources

About