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Perdeuteration of c...
Perdeuteration of cholesterol for neutron scattering applications using recombinant Pichia pastoris
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- Moulin, Martine (author)
- Institut Laue-Langevin, 71, Avenue des Martyrs, Grenoble 38042, France; Faculty of Natural Sciences, Keele University, Keele, Staffordshire ST5 5BG, United Kingdom,Inst Laue Langevin, Grenoble, France; Keele Univ, Fac Nat Sci, Staffs, England
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- Strohmeier, Gernot A. (author)
- acib, Austrian Centre of Industrial Biotechnology GmbH, 8010 Graz, Austria; Institute of Organic Chemistry, NAWI Graz, Graz University of Technology, 8010 Graz, Austria,Acib, Austrian Ctr Ind Biotechnol GmbH, Graz, Austria; Graz Univ Technol, Inst Organ Chem, NAWI Graz, Graz, Austria
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- Hirz, Melanie (author)
- Institute of Molecular Biotechnology, NAWI Graz, BioTechMed Graz, Graz University of Technology, 8010 Graz, Austria,Graz Univ Technol, Inst Mol Biotechnol, BioTechMed Graz, NAWI Graz, Graz, Austria
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- Thompson, Katherine C. (author)
- Department of Biological Sciences and Institute of Structural and Molecular Biology, Birkbeck College, University of London, Malet Street, London WC1E 7HX, United Kingdom,Univ London, Birkbeck Coll, Dept Biol Sci, London, England; Univ London, Birkbeck Coll, Inst Struct & Mol Biol, London, England
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- Rennie, Adrian R., 1957- (author)
- Uppsala universitet,Materialfysik,Centre for Neutron Scattering, Uppsala University, 751 20 Uppsala, Sweden
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- Campbell, Richard A. (author)
- Institut Laue-Langevin, 71, Avenue des Martyrs, Grenoble 38042, France,Inst Laue Langevin, Grenoble, France
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- Pichler, Harald (author)
- acib, Austrian Centre of Industrial Biotechnology GmbH, 8010 Graz, Austria; Institute of Molecular Biotechnology, NAWI Graz, BioTechMed Graz, Graz University of Technology, 8010 Graz, Austria,Acib, Austrian Ctr Ind Biotechnol GmbH, Graz, Austria; Graz Univ Technol, Inst Mol Biotechnol, BioTechMed Graz, NAWI Graz, Graz, Austria
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- Maric, Selma (author)
- Malmö universitet,Institutionen för biomedicinsk vetenskap (BMV),Biofilms Research Center for Biointerfaces,Malmö Univ, Fac Hlth & Soc, Biofilms Res Ctr Biointerfaces, Malmö, Sweden; Malmö Univ, Fac Hlth & Soc, Biomed Sci Dept, Malmö, Sweden
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- Forsyth, V. Trevor (author)
- Institut Laue-Langevin, 71, Avenue des Martyrs, Grenoble 38042, France; Faculty of Natural Sciences, Keele University, Keele, Staffordshire ST5 5BG, United Kingdom,Inst Laue Langevin, Grenoble, France; Keele Univ, Fac Nat Sci, Staffs, England
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- Haertlein, Michael (author)
- Institut Laue-Langevin, 71, Avenue des Martyrs, Grenoble 38042, France,Inst Laue Langevin, Grenoble, France
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(creator_code:org_t)
- Elsevier, 2018
- 2018
- English.
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In: Chemistry and Physics of Lipids. - : Elsevier. - 0009-3084 .- 1873-2941. ; 212, s. 80-87
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Abstract
Subject headings
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- Deuteration of biomolecules has a major impact on both quality and scope of neutron scattering experiments. Cholesterol is a major component of mammalian cells, where it plays a critical role in membrane permeability, rigidity and dynamics, and contributes to specific membrane structures such as lipid rafts. Cholesterol is the main cargo in low and high-density lipoprotein complexes (i.e. LDL, HDL) and is directly implicated in several pathogenic conditions such as coronary artery disease which leads to 17 million deaths annually. Neutron scattering studies on membranes or lipid-protein complexes exploiting contrast variation have been limited by the lack of availability of fully deuterated biomolecules and especially perdeuterated cholesterol. The availability of perdeuterated cholesterol provides a unique way of probing the structural and dynamical properties of the lipoprotein complexes that underly many of these disease conditions. Here we describe a procedure for in vivo production of perdeuterated recombinant cholesterol in lipid-engineered Pichia pastoris using flask and fed batch fermenter cultures in deuterated minimal medium. Perdeuteration of the purified cholesterol was verified by mass spectrometry and its use in a neutron scattering study was demonstrated by neutron reflectometry measurements using the FIGARO instrument at the ILL.
Subject headings
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Keyword
- Perdeuteration
- Cholesterol
- Pichia pastoris
- Neutron scattering
- Lipid engineering
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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Moulin, Martine
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Strohmeier, Gern ...
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Hirz, Melanie
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Thompson, Kather ...
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Rennie, Adrian R ...
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Campbell, Richar ...
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Pichler, Harald
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Maric, Selma
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Forsyth, V. Trev ...
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Haertlein, Micha ...
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- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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and Biochemistry and ...
- Articles in the publication
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Chemistry and Ph ...
- By the university
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Malmö University
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Uppsala University