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Engineering a Chemical Switch into the Light‐driven Proton Pump Proteorhodopsin by Cysteine Mutagenesis and Thiol Modification
Author(s) -
Harder Daniel,
Hirschi Stephan,
Ucurum Zöhre,
Goers Roland,
Meier Wolfgang,
Müller Daniel J.,
Fotiadis Dimitrios
Publication year - 2016
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201601537
Subject(s) - cysteine , mutagenesis , chemistry , residue (chemistry) , proton , electrochemical gradient , chemical modification , proton pump , combinatorial chemistry , nanotechnology , mutant , biophysics , membrane , biochemistry , materials science , biology , physics , atpase , quantum mechanics , gene , enzyme
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.