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Nanotechnology Reviews

Editor-in-Chief: Kumar, Challa

Ed. by Hamblin, Michael R. / Bianco, Alberto / Jin, Rongchao / Köhler, J. Michael / Hudait, Mantu K. / Dai, Ning / Lytton-Jean, Abigail / Xie, Jianping / Bryan, Lynn A. / Thiessen, Rose / Alexiou, Christoph / Lee, Jae-Seung / Delville, Marie-Helene / Yan, Ning / Baretzky, Brigitte / Burg, Thomas P. / Fenniri, Hicham / Yang, Jun / Hosmane, Narayan S. / Dufrene, Yves / Podila, Ramakrishna / Eswaramoorthy, Muthusamy

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Volume 6, Issue 1 (Feb 2017)

Issues

Proteoliposomes – a system to study membrane proteins under buffer gradients by cryo-EM

Kushal Sejwal
  • Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, Basel 4056, Switzerland
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Mohamed Chami
  • Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, Basel 4056, Switzerland
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Paul Baumgartner
  • Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, Basel 4056, Switzerland
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  • De Gruyter OnlineGoogle Scholar
/ Julia Kowal
  • Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, Basel 4056, Switzerland
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Shirley A. Müller
  • Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, Basel 4056, Switzerland
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Henning StahlbergORCID iD: http://orcid.org/0000-0002-1185-4592
Published Online: 2017-01-20 | DOI: https://doi.org/10.1515/ntrev-2016-0081

Abstract

Membrane proteins are vital to life and major therapeutic targets. Yet, understanding how they function is limited by a lack of structural information. In biological cells, membrane proteins reside in lipidic membranes and typically experience different buffer conditions on both sides of the membrane or even electric potentials and transmembrane gradients across the membranes. Proteoliposomes, which are lipidic vesicles filled with reconstituted membrane proteins, provide an ideal model system for structural and functional studies of membrane proteins under conditions that mimic nature to a certain degree. We discuss methods for the formation of liposomes and proteoliposomes, their imaging by cryo-electron microscopy, and the structural analysis of proteins present in their bilayer. We suggest the formation of ordered arrays akin to weakly ordered two-dimensional (2D) crystals in the bilayer of liposomes as a means to achieve high-resolution, and subsequent buffer modification as a method to capture snapshots of membrane proteins in action.

Keywords: buffer gradient; cryo-electron microscopy; image processing; membrane proteins; proteoliposomes

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About the article

Kushal Sejwal

Kushal Sejwal received his BS in Life Sciences from the University of Delhi and MS in Bioinformatics from Jamia Millia Islamia, New Delhi, India. He worked as a research associate at the Rudolf Virchow Center, University of Würzburg, in Germany for 2 years, where he was first introduced to the field of cryo-electron microscopy. In October 2012, he moved to Basel, Switzerland, to pursue a PhD in the laboratory of Prof. Henning Stahlberg at the C-CINA, Biozentrum, University of Basel. He worked on structural elucidation of macromolecular complexes by cryo-EM and method development using proteoliposomes. In August 2016, he finished his PhD and started his postdoctoral training in the laboratory of Prof. Michel Steinmetz at the Paul Scherrer Institute in Switzerland.

Mohamed Chami

Mohamed Chami obtained his undergraduate degree in biology and geology at the University of Oujda in Morocco. After receiving a master’s degree in biochemistry in 1994 from the University of Paris, France, he pursued a PhD on the structure of the cell envelope of Corynebacterium glutamicum at the same University. From 1999 to 2003, he worked as a post-doc on reconstitution and 2D crystallization of membrane proteins in the group of Jean-Louis Rigaud at the Institut Curie in Paris. He moved to Basel, Switzerland, in 2003 and worked until 2008 in the laboratory of Andreas Engel and, since 2009, in the laboratory of Henning Stahlberg as a senior scientist, where he is involved in the structure elucidation of membrane proteins using cryo-electron microscopy. Since January 2016, Dr. Chami had been directing the BioEM laboratory, the service facility for electron microscopy in the life sciences at the Biozentrum of the University of Basel, Switzerland.

Julia Kowal

Julia Kowal from Poland got her master’s degree in biochemistry at the Jagiellonian University, Kraków, Poland, and then obtained her PhD in biophysics in the group of Prof. Andreas Engel at the University of Basel, Switzerland. She then studied potassium channel structures by cryo-electron microscopy in the group of Prof. Stahlberg at C-CINA from 2011 to 2016. Since 2016, she has been a post-doc in the group of Prof. Locher at the ETH Zurich, Switzerland, where she continues studying membrane proteins by cryo-electron microscopy.

Shirley A. Müller

Shirley A. Müller received a PhD in Chemistry from the University of Exeter, UK, in 1977. Since then, the main focus of her scientific career has been mass measurements by scanning transmission electron microscopy, in the group of Andreas Engel at the Maurice E. Müller Institute for High-Resolution Electron Microscopy and at C-CINA (both at the Biozentrum, University of Basel, Switzerland). Her present tasks at C-CINA in the group of Henning Stahlberg are concerned with project management and scientific editing.

Henning Stahlberg

Henning Stahlberg studied physics at the Technical University of Berlin, Germany, from 1989 to 1992, and obtained his PhD from 1992 to 1997 in structural biology of membrane proteins at the EPFL in Lausanne, Switzerland, in the groups of Prof. Dubochet and Vogel. From 1998 to 2003, he studied membrane proteins by cryo-electron microscopy as a post-doc in the group of Prof. Engel at the Biozentrum, University of Basel, Switzerland. In 2003, he joined the University of California in Davis, CA, USA, as Assistant Professor, where he became tenured Associate Professor in 2007. Since 2009, he has been directing the Center for Cellular Imaging and NanoAnalytics (C-CINA) as Professor of the Biozentrum at the University of Basel, Switzerland.


Received: 2016-09-12

Accepted: 2016-12-01

Published Online: 2017-01-20

Published in Print: 2017-02-01


Citation Information: Nanotechnology Reviews, ISSN (Online) 2191-9097, ISSN (Print) 2191-9089, DOI: https://doi.org/10.1515/ntrev-2016-0081.

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