title: Variation of the detergent-binding capacity and phospholipid content of membrane proteins when purified in different detergents.
abstract: Purified membrane proteins are ternary complexes consisting of protein, lipid, and detergent. Information about the amounts of detergent and endogenous phospholipid molecules bound to purified membrane proteins is largely lacking. In this systematic study, three model membrane proteins of different oligomeric states were purified in nine different detergents at commonly used concentrations and characterized biochemically and biophysically. Detergent-binding capacities and phospholipid contents of the model proteins were determined and compared. The insights on ternary complexes obtained from the experimental results, when put into a general context, are summarized as follows. 1), The amount of detergent and 2) the amount of endogenous phospholipids bound to purified membrane proteins are dependent on the size of the hydrophobic lipid-accessible protein surface areas and the physicochemical properties of the detergents used. 3), The size of the detergent and lipid belt surrounding the hydrophobic lipid-accessible surface of purified membrane proteins can be tuned by the appropriate choice of detergent. 4), The detergents n-nonyl-β-D-glucopyranoside and Cymal-5 have exceptional delipidating effects on ternary complexes. 5), The types of endogenous phospholipids bound to membrane proteins can vary depending on the detergent used for solubilization and purification. 6), Furthermore, we demonstrate that size-exclusion chromatography can be a suitable method for estimating the molecular mass of ternary complexes. The findings presented suggest a strategy to control and tune the numbers of detergent and endogenous phospholipid molecules bound to membrane proteins. These two parameters are potentially important for the successul crystallization of membrane proteins for structure determination by crystallographic approaches.
authors.affiliation: Institute of Biochemistry and Molecular Medicine, and Swiss National Centre of Competence in Research (NCCR) TransCure, University of Bern, Bern, Switzerland.
authors.forename: Hüseyin
authors.initials: H
authors.lastname: Ilgü
authors.affiliation: Institute of Biochemistry and Molecular Medicine, and Swiss National Centre of Competence in Research (NCCR) TransCure, University of Bern, Bern, Switzerland.
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Jeckelmann
authors.affiliation: Institute of Biochemistry and Molecular Medicine, and Swiss National Centre of Competence in Research (NCCR) TransCure, University of Bern, Bern, Switzerland.
authors.forename: María Salomé
authors.initials: MS
authors.lastname: Gachet
authors.affiliation: Institute of Biochemistry and Molecular Medicine, and Swiss National Centre of Competence in Research (NCCR) TransCure, University of Bern, Bern, Switzerland.
authors.forename: Rajendra
authors.initials: R
authors.lastname: Boggavarapu
authors.affiliation: Institute of Biochemistry and Molecular Medicine, and Swiss National Centre of Competence in Research (NCCR) TransCure, University of Bern, Bern, Switzerland.
authors.forename: Zöhre
authors.initials: Z
authors.lastname: Ucurum
authors.affiliation: Institute of Biochemistry and Molecular Medicine, and Swiss National Centre of Competence in Research (NCCR) TransCure, University of Bern, Bern, Switzerland.
authors.forename: Jürg
authors.initials: J
authors.lastname: Gertsch
authors.affiliation: Institute of Biochemistry and Molecular Medicine, and Swiss National Centre of Competence in Research (NCCR) TransCure, University of Bern, Bern, Switzerland. Electronic address: dimitrios.fotiadis@ibmm.unibe.ch.
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Fotiadis
mesh_terms: D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism; D002850:Chromatography, Gel; D016969:Desulfovibrio vulgaris / Q000378:metabolism; D003902:Detergents / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000378:metabolism; D008958:Models, Molecular; D008970:Molecular Weight; D010714:Phosphatidylethanolamines / Q000378:metabolism; D010715:Phosphatidylglycerols / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism*; D011485:Protein Binding; D000096982:Urea Transporters
chemical_list: D001426:Bacterial Proteins; D003902:Detergents; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D010714:Phosphatidylethanolamines; D010715:Phosphatidylglycerols; D010743:Phospholipids; D000096982:Urea Transporters; C483858:phosphatidylethanolamine
references.citation: Curr Opin Struct Biol. 1997 Oct;7(5):697-701
references.pmid: 9345629
references.citation: PLoS One. 2007 Sep 12;2(9):e880
references.pmid: 17849009
references.citation: Bioinformatics. 2006 Mar 1;22(5):623-5
references.pmid: 16397007
references.citation: J Biol Chem. 2007 Feb 2;282(5):2832-9
references.pmid: 17158458
references.citation: Nature. 2010 Sep 9;467(7312):233-6
references.pmid: 20829798
references.citation: J Bioenerg Biomembr. 1996 Feb;28(1):13-27
references.pmid: 8786233
references.citation: Biochim Biophys Acta. 2004 Nov 3;1666(1-2):105-17
references.pmid: 15519311
references.citation: Rapid Commun Mass Spectrom. 2007;21(11):1721-8
references.pmid: 17477452
references.citation: Protein Sci. 2003 Dec;12(12):2748-56
references.pmid: 14627735
references.citation: Nature. 2011 Apr 14;472(7342):243-6
references.pmid: 21423164
references.citation: Nature. 2013 Apr 25;496(7446):533-6
references.pmid: 23542591
references.citation: Can J Biochem Physiol. 1959 Aug;37(8):911-7
references.pmid: 13671378
references.citation: J Struct Biol. 2002 Mar;137(3):322-32
references.pmid: 12096900
references.citation: Proc Natl Acad Sci U S A. 2007 Sep 25;104(39):15294-8
references.pmid: 17881559
references.citation: Biochim Biophys Acta. 2000 Nov 23;1508(1-2):86-111
references.pmid: 11090820
references.citation: Nature. 2009 Mar 5;458(7234):47-52
references.pmid: 19262666
references.citation: Methods. 2008 Oct;46(2):73-82
references.pmid: 18625320
references.citation: Science. 2009 Jun 19;324(5934):1565-8
references.pmid: 19478139
references.citation: Anal Chem. 2010 Aug 1;82(15):6687-6694
references.pmid: 20698582
references.citation: J Am Chem Soc. 2009 Jun 3;131(21):7320-6
references.pmid: 19425578
references.citation: J Comput Chem. 2009 Oct;30(13):2157-64
references.pmid: 19229944
references.citation: Methods. 2008 Oct;46(2):62-72
references.pmid: 18952172
references.citation: J Struct Biol. 1992 Nov-Dec;109(3):219-34
references.pmid: 1296757
references.citation: Biochemistry. 2007 May 1;46(17):5200-8
references.pmid: 17407265
references.citation: Biophys J. 2011 Jan 19;100(2):L11-3
references.pmid: 21244820
references.citation: PLoS One. 2013 May 08;8(5):e62488
references.pmid: 23667481
references.citation: Micron. 2001 Jan;32(1):75-90
references.pmid: 10900383
references.citation: Biophys J. 2009 Jul 8;97(1):50-8
references.pmid: 19580743
references.citation: J Mol Biol. 1999 Jul 2;290(1):253-66
references.pmid: 10388571
references.citation: Nature. 2010 Oct 7;467(7316):734-8
references.pmid: 20877283
references.citation: Methods. 2007 Apr;41(4):388-97
references.pmid: 17367711
references.citation: Anal Biochem. 1985 Oct;150(1):76-85
references.pmid: 3843705
references.citation: Biochim Biophys Acta. 2012 May;1818(5):1351-8
references.pmid: 22285740
references.citation: Mass Spectrom Rev. 2003 Sep-Oct;22(5):332-64
references.pmid: 12949918
references.citation: Biophys J. 2006 Jan 1;90(1):310-7
references.pmid: 16214861
references.citation: Proc Natl Acad Sci U S A. 2007 Jun 26;104(26):10830-4
references.pmid: 17566106
references.citation: Nanotechnology. 2008 Sep 24;19(38):384014
references.pmid: 21832573
references.citation: Nature. 2009 Dec 10;462(7274):757-61
references.pmid: 19865084