title: Physicochemical factors controlling the activity and energy coupling of an ionic strength-gated ATP-binding cassette (ABC) transporter.
abstract: Cells control their volume through the accumulation of compatible solutes. The bacterial ATP-binding cassette transporter OpuA couples compatible solute uptake to ATP hydrolysis. Here, we study the gating mechanism and energy coupling of OpuA reconstituted in lipid nanodiscs. We show that anionic lipids are essential both for the gating and the energy coupling. The tight coupling between substrate binding on extracellular domains and ATP hydrolysis by cytoplasmic nucleotide-binding domains allows the study of transmembrane signaling in nanodiscs. From the tight coupling between processes at opposite sides of the membrane, we infer that the ATPase activity of OpuA in nanodiscs reflects solute translocation. Intriguingly, the substrate-dependent, ionic strength-gated ATPase activity of OpuA in nanodiscs is at least an order of magnitude higher than in lipid vesicles (i.e. with identical membrane lipid composition, ionic strength, and nucleotide and substrate concentrations). Even with the chemical components the same, the lateral pressure (profile) of the nanodiscs will differ from that of the vesicles. We thus propose that membrane tension limits translocation in vesicular systems. Increased macromolecular crowding does not activate OpuA but acts synergistically with ionic strength, presumably by favoring gating interactions of like-charged surfaces via excluded volume effects.
authors: From the Departments of Biochemistry and Biophysical Chemistry, Groningen Biomolecular Sciences and Biotechnology Institute, Netherlands Proteomics Centre.
authors: Akira
authors: A
authors: Karasawa
authors: Lotteke J Y M
authors: LJ
authors: Swier
authors: Marc C A
authors: MC
authors: Stuart
authors: Jos
authors: J
authors: Brouwers
authors: Bernd
authors: B
authors: Helms
authors: Bert
authors: B
authors: Poolman
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001692:Biological Transport; D055598:Chemical Phenomena; D002853:Chromatography, Liquid; D004735:Energy Transfer; D006868:Hydrolysis; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry; D008563:Membrane Lipids / Q000737:chemistry / Q000378:metabolism; D049329:Nanostructures / Q000737:chemistry; D009994:Osmolar Concentration; D011510:Proteolipids / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D008051:Lipid Bilayers; D008081:Liposomes; D008563:Membrane Lipids; D011510:Proteolipids; C020485:proteoliposomes; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; C412743:OpuA protein, Lactococcus lactis
keywords: ABC Transporter; BC Transporter; Cell Volume Regulation; Energetics; Gating; Gating Mechanism; Ionic Strength; Macromolecular Crowding; Membrane; Membrane Reconstitution; Membrane Transport
references: EMBO J. 2001 Oct 1;20(19):5412-20
references: 11574473
references: Biochemistry. 2003 Jan 21;42(2):410-20
references: 12525168
references: PLoS One. 2010 Apr 29;5(4):e10361
references: 20454456
references: Biochemistry. 2012 Dec 11;51(49):9782-95
references: 23163284
references: Protein Sci. 2004 Jan;13(1):125-33
references: 14691228
references: Methods Mol Biol. 2001;168:17-36
references: 11357625
references: J Biol Chem. 2003 Aug 8;278(32):29546-51
references: 12766159
references: Nature. 2007 Mar 8;446(7132):213-6
references: 17322901
references: Langmuir. 2012 Jun 26;28(25):9475-86
references: 22646139
references: EMBO J. 2010 Dec 1;29(23):3911-23
references: 21037553
references: Nature. 2012 Oct 4;490(7418):126-30
references: 22940865
references: J Biol Chem. 2011 Nov 11;286(45):39489-96
references: 21937435
references: FEBS Lett. 2011 Nov 16;585(22):3533-7
references: 22020218
references: Biochemistry. 1988 Feb 9;27(3):865-72
references: 3284574
references: Biochemistry. 2010 Oct 19;49(41):8847-56
references: 20828170
references: J Am Chem Soc. 2011 Nov 16;133(45):18318-27
references: 21951206
references: Methods Enzymol. 2009;464:211-31
references: 19903557
references: FEBS Lett. 2010 May 3;584(9):1721-7
references: 19836392
references: PLoS One. 2012;7(4):e34836
references: 22529943
references: Biochemistry. 2005 Dec 13;44(49):16301-9
references: 16331991
references: Biophys J. 1997 Mar;72(3):1022-30
references: 9138553
references: Biophys J. 2007 Dec 1;93(11):3884-99
references: 17704167
references: J Biol Chem. 2006 Oct 6;281(40):29830-9
references: 16844687
references: Mol Microbiol. 2010 Oct;78(1):13-34
references: 20923416
references: J Biol Chem. 2012 Aug 10;287(33):27813-22
references: 22740696
references: Proc Natl Acad Sci U S A. 2006 Jul 11;103(28):10624-9
references: 16815971
references: Trends Biochem Sci. 2005 Oct;30(10):536-41
references: 16125938
references: J Biol Chem. 2007 Mar 2;282(9):6556-63
references: 17200119
references: Biochemistry. 2008 Aug 5;47(31):8176-85
references: 18620422
references: Proc Natl Acad Sci U S A. 2006 Aug 1;103(31):11509-14
references: 16864771
references: J Biol Chem. 2005 Dec 16;280(50):41387-94
references: 16239220
references: J Biol Chem. 1995 Jul 7;270(27):16167-75
references: 7608182
references: Proc Natl Acad Sci U S A. 2000 Apr 11;97(8):3987-92
references: 10760270
references: Biochim Biophys Acta. 2013 Apr;1828(4):1222-9
references: 23276833
references: Biochemistry. 2001 Jun 19;40(24):7324-33
references: 11401581
references: Arch Biochem Biophys. 2006 Jun 15;450(2):215-22
references: 16620766
references: EMBO J. 2011 Apr 6;30(7):1221-9
references: 21364531
references: Biochim Biophys Acta. 2012 Mar;1818(3):359-66
references: 22051018
references: J Biol Chem. 2009 Mar 20;284(12):7897-902
references: 19139097
references: Proc Natl Acad Sci U S A. 2011 Jun 7;108(23):9390-5
references: 21606342
references: J Am Chem Soc. 2010 Jul 21;132(28):9513-5
references: 20578693
references: Nat Protoc. 2008;3(2):256-66
references: 18274528
references: J Mol Biol. 2012 Mar 16;417(1-2):95-111
references: 22306739
references: EMBO J. 2007 Apr 18;26(8):1995-2004
references: 17396152
references: Proc Natl Acad Sci U S A. 2012 Mar 13;109(11):4104-9
references: 22378651
references: Comp Biochem Physiol A Mol Integr Physiol. 2001 Oct;130(3):437-60
references: 11913457
references: J Biol Chem. 2011 Oct 28;286(43):37280-91
references: 21878634
references: J Biol Chem. 2011 Jan 14;286(2):1420-8
references: 20966068
references: Proc Natl Acad Sci U S A. 2007 May 1;104(18):7682-7
references: 17452637
references: EMBO J. 2001 Dec 17;20(24):7022-32
references: 11742979