Experimental platform for the functional investigation of membrane proteins in giant unilamellar vesicles.

Dolder, Nicolas; Müller, Philipp; von Ballmoos, Christoph (2022). Experimental platform for the functional investigation of membrane proteins in giant unilamellar vesicles. Soft matter, 18(31), pp. 5877-5893. Royal Society of Chemistry 10.1039/d2sm00551d

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Giant unilamellar vesicles (GUVs) are micrometer-sized model membrane systems that can be viewed directly under the microscope. They serve as scaffolds for the bottom-up creation of synthetic cells, targeted drug delivery and have been widely used to study membrane related phenomena in vitro. GUVs are also of interest for the functional investigation of membrane proteins that carry out many key cellular functions. A major hurdle to a wider application of GUVs in this field is the diversity of existing protocols that are optimized for individual proteins. Here, we compare PVA assisted and electroformation techniques for GUV formation under physiologically relevant conditions, and analyze the effect of immobilization on vesicle structure and membrane tightness towards small substrates and protons. There, differences in terms of yield, size, and leakage of GUVs produced by PVA assisted swelling and electroformation were found, dependent on salt and buffer composition. Using fusion of oppositely charged membranes to reconstitute a model membrane protein, we find that empty vesicles and proteoliposomes show similar fusion behavior, which allows for a rapid estimation of protein incorporation using fluorescent lipids.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Department of Chemistry, Biochemistry and Pharmaceutical Sciences (DCBP)

UniBE Contributor:

Dolder, Nicolas, Müller, Philipp Isaak, von Ballmoos, Christoph

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry
000 Computer science, knowledge & systems

ISSN:

1744-683X

Publisher:

Royal Society of Chemistry

Language:

English

Submitter:

Pubmed Import

Date Deposited:

03 Aug 2022 09:42

Last Modified:

05 Dec 2022 16:22

Publisher DOI:

10.1039/d2sm00551d

PubMed ID:

35916307

BORIS DOI:

10.48350/171711

URI:

https://boris.unibe.ch/id/eprint/171711

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