caa a22 4500 60549813X CHVBK 20210128100544.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00253-014-6082-z doi (NATIONALLICENCE)springer-10.1007/s00253-014-6082-z Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli [Elektronische Daten] [Selma Maric, Mikkel Thygesen, Jürgen Schiller, Magdalena Marek, Martine Moulin, Michael Haertlein, V. Forsyth, Mikhail Bogdanov, William Dowhan, Lise Arleth, Thomas Pomorski] Phosphatidylcholine (PC) is a major component of eukaryotic cell membranes and one of the most commonly used phospholipids for reconstitution of membrane proteins into carrier systems such as lipid vesicles, micelles and nanodiscs. Selectively deuterated versions of this lipid have many applications, especially in structural studies using techniques such as NMR, neutron reflectivity and small-angle neutron scattering. Here we present a comprehensive study of selective deuteration of phosphatidylcholine through biosynthesis in a genetically modified strain of Escherichia coli. By carefully tuning the deuteration level in E. coli growth media and varying the deuteration of supplemented carbon sources, we show that it is possible to achieve a controlled deuteration for three distinct parts of the PC lipid molecule, namely the (a) lipid head group, (b) glycerol backbone and (c) fatty acyl tail. This biosynthetic approach paves the way for the synthesis of specifically deuterated, physiologically relevant phospholipid species which remain difficult to obtain through standard chemical synthesis. Springer-Verlag Berlin Heidelberg, 2014 Glycerophospholipids nationallicence Phosphatidylcholine nationallicence Selective deuteration nationallicence Biosynthesis nationallicence E. coli nationallicence Neutron scattering nationallicence NMR nationallicence Mass spectrometry nationallicence Evolution in the test tube nationallicence Maric Selma Structural Biophysics, Niels Bohr Institute, Faculty of Science, University of Copenhagen, Universitetsparken 5, 2100, Copenhagen, Denmark aut Thygesen Mikkel CARB Centre, Department of Chemistry, Faculty of Science, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark aut Schiller Jürgen Institut für Medizinische Physik und Biophysik, Medizinische Fakultät, Universität Leipzig, Härtelstrasse 16-18, 04107, Leipzig, Germany aut Marek Magdalena Center for Membrane Pumps in Cells and Disease, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark aut Moulin Martine Life Sciences Group, Institut Laue Langevin, 6 rue Jules Horowitz, CEDEX 9, BP156, 38042, Grenoble, France aut Haertlein Michael Life Sciences Group, Institut Laue Langevin, 6 rue Jules Horowitz, CEDEX 9, BP156, 38042, Grenoble, France aut Forsyth V. Life Sciences Group, Institut Laue Langevin, 6 rue Jules Horowitz, CEDEX 9, BP156, 38042, Grenoble, France aut Bogdanov Mikhail Department of Biochemistry and Molecular Biology, University of Texas Medical School at Houston, 77030, Houston, TX, USA aut Dowhan William Department of Biochemistry and Molecular Biology, University of Texas Medical School at Houston, 77030, Houston, TX, USA aut Arleth Lise Structural Biophysics, Niels Bohr Institute, Faculty of Science, University of Copenhagen, Universitetsparken 5, 2100, Copenhagen, Denmark aut Pomorski Thomas Center for Membrane Pumps in Cells and Disease, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/1(2015-01-01), 241-254 0175-7598 99:1<241 2015 99 253 https://doi.org/10.1007/s00253-014-6082-z text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6082-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Maric Selma Structural Biophysics, Niels Bohr Institute, Faculty of Science, University of Copenhagen, Universitetsparken 5, 2100, Copenhagen, Denmark aut NATIONALLICENCE 700 1- Thygesen Mikkel CARB Centre, Department of Chemistry, Faculty of Science, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark aut NATIONALLICENCE 700 1- Schiller Jürgen Institut für Medizinische Physik und Biophysik, Medizinische Fakultät, Universität Leipzig, Härtelstrasse 16-18, 04107, Leipzig, Germany aut NATIONALLICENCE 700 1- Marek Magdalena Center for Membrane Pumps in Cells and Disease, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark aut NATIONALLICENCE 700 1- Moulin Martine Life Sciences Group, Institut Laue Langevin, 6 rue Jules Horowitz, CEDEX 9, BP156, 38042, Grenoble, France aut NATIONALLICENCE 700 1- Haertlein Michael Life Sciences Group, Institut Laue Langevin, 6 rue Jules Horowitz, CEDEX 9, BP156, 38042, Grenoble, France aut NATIONALLICENCE 700 1- Forsyth V. Life Sciences Group, Institut Laue Langevin, 6 rue Jules Horowitz, CEDEX 9, BP156, 38042, Grenoble, France aut NATIONALLICENCE 700 1- Bogdanov Mikhail Department of Biochemistry and Molecular Biology, University of Texas Medical School at Houston, 77030, Houston, TX, USA aut NATIONALLICENCE 700 1- Dowhan William Department of Biochemistry and Molecular Biology, University of Texas Medical School at Houston, 77030, Houston, TX, USA aut NATIONALLICENCE 700 1- Arleth Lise Structural Biophysics, Niels Bohr Institute, Faculty of Science, University of Copenhagen, Universitetsparken 5, 2100, Copenhagen, Denmark aut NATIONALLICENCE 700 1- Pomorski Thomas Center for Membrane Pumps in Cells and Disease, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/1(2015-01-01), 241-254 0175-7598 99:1<241 2015 99 253