Deutschmann, Sabina; Rimle, Lukas; von Ballmoos, Christoph (2022). Rapid Estimation of Membrane Protein Orientation in Liposomes. ChemBioChem, 23(2), e202100543. Wiley-VCH 10.1002/cbic.202100543
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The topological organization of proteins embedded in biological membranes is crucial for the tight interplay between these enzymes and their accessibility to substrates in order to fulfil enzymatic activity. The orientation of a membrane protein reconstituted in artificial membranes depends on many parameters and is hardly predictable. Here, we present a convenient approach to assess this important property independent of the enzymatic activity of the reconstituted protein. Based on cysteine-specific chemical modification of a target membrane protein with a cyanine fluorophore and a corresponding membrane-impermeable fluorescence quencher, the novel strategy allows rapid evaluation of the distribution of the two orientations after reconstitution. The assay has been tested for the respiratory complexes bo3 oxidase and ATP synthase of Escherichia coli and the results agree well with other orientation determination approaches. Given the simple procedure, the proposed method is a powerful tool for optimization of reconstitution conditions or quantitative orientation information prior to functional measurements.
Item Type: |
Journal Article (Original Article) |
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Division/Institute: |
08 Faculty of Science > Department of Chemistry, Biochemistry and Pharmaceutical Sciences (DCBP) |
Graduate School: |
Graduate School for Cellular and Biomedical Sciences (GCB) |
UniBE Contributor: |
Deutschmann, Sabina Barbara, Rimle, Lukas, von Ballmoos, Christoph |
Subjects: |
500 Science > 570 Life sciences; biology 500 Science > 540 Chemistry |
ISSN: |
1439-4227 |
Publisher: |
Wiley-VCH |
Language: |
English |
Submitter: |
Lukas Rimle |
Date Deposited: |
03 Jan 2022 11:12 |
Last Modified: |
19 Jul 2023 08:15 |
Publisher DOI: |
10.1002/cbic.202100543 |
PubMed ID: |
34763366 |
Uncontrolled Keywords: |
fluorescence quenching liposomes membrane proteins orientation determination site-specific modification of proteins |
BORIS DOI: |
10.48350/162078 |
URI: |
https://boris.unibe.ch/id/eprint/162078 |
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