Next Article in Journal
Three-Dimensional Radiographic Evaluation of the Malar Bone Engagement Available for Ideal Zygomatic Implant Placement
Previous Article in Journal
Multiplex Cell Fate Tracking by Flow Cytometry
Protocol

Engineering and Production of the Light-Driven Proton Pump Bacteriorhodopsin in 2D Crystals for Basic Research and Applied Technologies

1
Institute of Biochemistry and Molecular Medicine, and Swiss National Centre of Competence in Research (NCCR) TransCure, University of Bern, 3012 Bern, Switzerland
2
Department of Physics, Genetic Biophysics, Freie Universität Berlin, 14195 Berlin, Germany
*
Authors to whom correspondence should be addressed.
Methods Protoc. 2020, 3(3), 51; https://doi.org/10.3390/mps3030051
Received: 6 July 2020 / Revised: 18 July 2020 / Accepted: 19 July 2020 / Published: 22 July 2020
The light-driven proton pump bacteriorhodopsin (BR) from the extreme halophilic archaeon Halobacterium salinarum is a retinal-binding protein, which forms highly ordered and thermally stable 2D crystals in native membranes (termed purple membranes). BR and purple membranes (PMs) have been and are still being intensively studied by numerous researchers from different scientific disciplines. Furthermore, PMs are being successfully used in new, emerging technologies such as bioelectronics and bionanotechnology. Most published studies used the wild-type form of BR, because of the intrinsic difficulty to produce genetically modified versions in purple membranes homologously. However, modification and engineering is crucial for studies in basic research and, in particular, to tailor BR for specific applications in applied sciences. We present an extensive and detailed protocol ranging from the genetic modification and cultivation of H. salinarum to the isolation, and biochemical, biophysical and functional characterization of BR and purple membranes. Pitfalls and problems of the homologous expression of BR versions in H. salinarum are discussed and possible solutions presented. The protocol is intended to facilitate the access to genetically modified BR versions for researchers of different scientific disciplines, thus increasing the application of this versatile biomaterial. View Full-Text
Keywords: bacteriorhodopsin; bioelectronics; biomaterial; bionanotechnology; Halobacterium salinarum; light-driven proton pump; protein engineering; purple membranes bacteriorhodopsin; bioelectronics; biomaterial; bionanotechnology; Halobacterium salinarum; light-driven proton pump; protein engineering; purple membranes
Show Figures

Figure 1

MDPI and ACS Style

Stauffer, M.; Hirschi, S.; Ucurum, Z.; Harder, D.; Schlesinger, R.; Fotiadis, D. Engineering and Production of the Light-Driven Proton Pump Bacteriorhodopsin in 2D Crystals for Basic Research and Applied Technologies. Methods Protoc. 2020, 3, 51. https://doi.org/10.3390/mps3030051

AMA Style

Stauffer M, Hirschi S, Ucurum Z, Harder D, Schlesinger R, Fotiadis D. Engineering and Production of the Light-Driven Proton Pump Bacteriorhodopsin in 2D Crystals for Basic Research and Applied Technologies. Methods and Protocols. 2020; 3(3):51. https://doi.org/10.3390/mps3030051

Chicago/Turabian Style

Stauffer, Mirko, Stephan Hirschi, Zöhre Ucurum, Daniel Harder, Ramona Schlesinger, and Dimitrios Fotiadis. 2020. "Engineering and Production of the Light-Driven Proton Pump Bacteriorhodopsin in 2D Crystals for Basic Research and Applied Technologies" Methods and Protocols 3, no. 3: 51. https://doi.org/10.3390/mps3030051

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop