Bifunctional DNA Duplexes Permit Efficient Incorporation of pH Probes into Liposomes.

Dolder, Nicolas; von Ballmoos, Christoph (2020). Bifunctional DNA Duplexes Permit Efficient Incorporation of pH Probes into Liposomes. ChemBioChem, 21(15), pp. 2219-2224. Wiley-VCH 10.1002/cbic.202000146

[img] Text
manuscript_accepted article_dolder.pdf - Published Version
Restricted to registered users only
Available under License Publisher holds Copyright.

Download (995kB) | Request a copy
[img] Text
cbic.202000146.pdf - Published Version
Restricted to registered users only
Available under License Publisher holds Copyright.

Download (889kB) | Request a copy

Enzyme-mediated proton transport across biological membranes is critical for many vital cellular processes. pH-sensitive fluorescent dyes are an indispensable tool for investigating the molecular mechanism of proton-translocating enzymes. Here, we present a novel strategy to entrap pH-sensitive probes in the lumen of liposomes that has several advantages over the use of soluble or lipid-coupled probes. In our approach, the pH sensor is linked to a DNA oligomer with a sequence complementary to a second oligomer modified with a lipophilic moiety that anchors the DNA conjugate to the inner and outer leaflets of the lipid bilayer. The use of DNA as a scaffold allows subsequent selective enzymatic removal of the probe in the outer bilayer leaflet. The method shows a high yield of insertion and is compatible with reconstitution of membrane proteins by different methods. The usefulness of the conjugate for time-resolved proton pumping measurements was demonstrated by using two large membrane protein complexes.

Item Type:

Journal Article (Original Article)

Division/Institute:

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

UniBE Contributor:

Dolder, Nicolas and von Ballmoos, Christoph

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry

ISSN:

1439-4227

Publisher:

Wiley-VCH

Language:

English

Submitter:

Christoph von Ballmoos

Date Deposited:

03 Nov 2020 09:25

Last Modified:

03 Nov 2020 09:34

Publisher DOI:

10.1002/cbic.202000146

PubMed ID:

32181556

Uncontrolled Keywords:

DNA conjugates carboxyfluorescein liposomes membrane proteins pH sensors

BORIS DOI:

10.7892/boris.147125

URI:

https://boris.unibe.ch/id/eprint/147125

Actions (login required)

Edit item Edit item
Provide Feedback