Engineering a Chemical Switch into the Light-driven Proton Pump Proteorhodopsin by Cysteine Mutagenesis and Thiol Modification.

Harder, Daniel; Hirschi, Stephan; Ucurum Fotiadis, Zöhre; Goers, Roland; Meier, Wolfgang; Müller, Daniel J; Fotiadis, Dimitrios José (2016). Engineering a Chemical Switch into the Light-driven Proton Pump Proteorhodopsin by Cysteine Mutagenesis and Thiol Modification. Angewandte Chemie (International ed.), 55(31), pp. 8846-8849. Wiley-VCH 10.1002/anie.201601537

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For applications in synthetic biology, for example, the bottom-up assembly of biomolecular nanofactories, modules of specific and controllable functionalities are essential. Of fundamental importance in such systems are energizing modules, which are able to establish an electrochemical gradient across a vesicular membrane as an energy source for powering other modules. Light-driven proton pumps like proteorhodopsin (PR) are excellent candidates for efficient energy conversion. We have extended the versatility of PR by implementing an on/off switch based on reversible chemical modification of a site-specifically introduced cysteine residue. The position of this cysteine residue in PR was identified by structure-based cysteine mutagenesis combined with a proton-pumping assay using E. coli cells overexpressing PR and PR proteoliposomes. The identified PR mutant represents the first light-driven proton pump that can be chemically switched on/off depending on the requirements of the molecular system.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Biochemistry and Molecular Medicine

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Harder, Daniel; Hirschi, Stephan; Ucurum Fotiadis, Zöhre and Fotiadis, Dimitrios José

Subjects:

500 Science > 570 Life sciences; biology
600 Technology > 610 Medicine & health

ISSN:

1433-7851

Publisher:

Wiley-VCH

Language:

English

Submitter:

Barbara Järmann-Bangerter

Date Deposited:

19 Apr 2017 10:35

Last Modified:

19 Apr 2017 10:35

Publisher DOI:

10.1002/anie.201601537

PubMed ID:

27294681

Uncontrolled Keywords:

light-driven proton pumps; membrane proteins; protein engineering; proteorhodopsin; synthetic biology

BORIS DOI:

10.7892/boris.94011

URI:

https://boris.unibe.ch/id/eprint/94011

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