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Controlled delivery of membrane proteins to artificial lipid bilayers by nystatin-ergosterol modulated vesicle fusion

Controlled delivery of membrane proteins to artificial lipid bilayers by nystatin-ergosterol modulated vesicle fusion
Controlled delivery of membrane proteins to artificial lipid bilayers by nystatin-ergosterol modulated vesicle fusion
The study of ion channels and other membrane proteins and their potential use as biosensors and drug screening targets require their reconstitution in an artificial membrane. These applications would greatly benefit from microfabricated devices in which stable artificial lipid bilayers can be rapidly and reliably formed. However, the amount of protein delivered to the bilayer must be carefully controlled. A vesicle fusion technique is investigated where composite ion channels of the polyene antibiotic nystatin and the sterol ergosterol are employed to render protein-carrying vesicles fusogenic After fusion with an ergosterol-free artificial bilayer the nystatin-ergosterol channels do not dissociate immediately and thus cause a transient current signal that marks the vesicle fusion event. Experimental pitfalls of this method were identified, the influence of the nystatin and ergosterol concentration on the fusion rate and the shape of the fusion event marker was explored, and the number of different lipid was reduced. Under these conditions, the B-amyloid peptide could be delivered in a controlled manner to a standard planar bilayer. Additionally, the electrical recordings were obtained of vesicles fusing with a planar lipid bilayer in a microfabricated device, demonstrating the suitability of nystatin-ergosterol modulated vesicle fusion for protein delivery within microsystems.
21-30
de Planque, M R R
9d9c48a6-2a14-48fd-baea-54cfbd6aceeb
Mendes, G P
e5ce3fa2-6551-47ae-bd52-65aecbbc01ba
Zagnoni, M
b5c46de0-013d-470e-adc8-95ce8ad2545d
Sandison, M E
18641143-283c-4a25-a4d1-a46453b7178e
Fisher, K H
9718c40c-3083-4836-b697-239523b85917
Berry, R M
102975a1-c424-417c-82d1-42a5740df078
Watts, A
e234f61d-ba7f-4ef8-a632-9fec7c896c5b
Morgan, H
de00d59f-a5a2-48c4-a99a-1d5dd7854174
de Planque, M R R
9d9c48a6-2a14-48fd-baea-54cfbd6aceeb
Mendes, G P
e5ce3fa2-6551-47ae-bd52-65aecbbc01ba
Zagnoni, M
b5c46de0-013d-470e-adc8-95ce8ad2545d
Sandison, M E
18641143-283c-4a25-a4d1-a46453b7178e
Fisher, K H
9718c40c-3083-4836-b697-239523b85917
Berry, R M
102975a1-c424-417c-82d1-42a5740df078
Watts, A
e234f61d-ba7f-4ef8-a632-9fec7c896c5b
Morgan, H
de00d59f-a5a2-48c4-a99a-1d5dd7854174

de Planque, M R R, Mendes, G P, Zagnoni, M, Sandison, M E, Fisher, K H, Berry, R M, Watts, A and Morgan, H (2006) Controlled delivery of membrane proteins to artificial lipid bilayers by nystatin-ergosterol modulated vesicle fusion. IEE Proceedings - Nanobiotechnology, 153 (2), 21-30.

Record type: Article

Abstract

The study of ion channels and other membrane proteins and their potential use as biosensors and drug screening targets require their reconstitution in an artificial membrane. These applications would greatly benefit from microfabricated devices in which stable artificial lipid bilayers can be rapidly and reliably formed. However, the amount of protein delivered to the bilayer must be carefully controlled. A vesicle fusion technique is investigated where composite ion channels of the polyene antibiotic nystatin and the sterol ergosterol are employed to render protein-carrying vesicles fusogenic After fusion with an ergosterol-free artificial bilayer the nystatin-ergosterol channels do not dissociate immediately and thus cause a transient current signal that marks the vesicle fusion event. Experimental pitfalls of this method were identified, the influence of the nystatin and ergosterol concentration on the fusion rate and the shape of the fusion event marker was explored, and the number of different lipid was reduced. Under these conditions, the B-amyloid peptide could be delivered in a controlled manner to a standard planar bilayer. Additionally, the electrical recordings were obtained of vesicles fusing with a planar lipid bilayer in a microfabricated device, demonstrating the suitability of nystatin-ergosterol modulated vesicle fusion for protein delivery within microsystems.

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dePlanque_IEEProcNanobiotechnol_2006.pdf - Other
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More information

Published date: April 2006
Organisations: Nanoelectronics and Nanotechnology

Identifiers

Local EPrints ID: 263178
URI: https://eprints.soton.ac.uk/id/eprint/263178
PURE UUID: 1a1edaa4-d7fb-4a5b-ab9f-35fbfe40fea5
ORCID for H Morgan: ORCID iD orcid.org/0000-0003-4850-5676

Catalogue record

Date deposited: 14 Nov 2006
Last modified: 17 Sep 2019 00:52

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