title: F1-ATPase of Escherichia coli: the ε- inhibited state forms after ATP hydrolysis, is distinct from the ADP-inhibited state, and responds dynamically to catalytic site ligands.
abstract: F1-ATPase is the catalytic complex of rotary nanomotor ATP synthases. Bacterial ATP synthases can be autoinhibited by the C-terminal domain of subunit ε, which partially inserts into the enzyme's central rotor cavity to block functional subunit rotation. Using a kinetic, optical assay of F1·ε binding and dissociation, we show that formation of the extended, inhibitory conformation of ε (εX) initiates after ATP hydrolysis at the catalytic dwell step. Prehydrolysis conditions prevent formation of the εX state, and post-hydrolysis conditions stabilize it. We also show that ε inhibition and ADP inhibition are distinct, competing processes that can follow the catalytic dwell. We show that the N-terminal domain of ε is responsible for initial binding to F1 and provides most of the binding energy. Without the C-terminal domain, partial inhibition by the ε N-terminal domain is due to enhanced ADP inhibition. The rapid effects of catalytic site ligands on conformational changes of F1-bound ε suggest dynamic conformational and rotational mobility in F1 that is paused near the catalytic dwell position.
authors: Department of Biochemistry and Molecular Biology, State University of New York Upstate Medical University, Syracuse, NY 13210, USA.
authors: Naman B
authors: NB
authors: Shah
authors: Marcus L
authors: ML
authors: Hutcheon
authors: Brian K
authors: BK
authors: Haarer
authors: Thomas M
authors: TM
authors: Duncan
mesh_terms: D000244:Adenosine Diphosphate / Q000737:chemistry*; D000255:Adenosine Triphosphate / Q000737:chemistry*; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001692:Biological Transport; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D004926:Escherichia coli / Q000378:metabolism*; D006868:Hydrolysis; D007700:Kinetics; D008024:Ligands; D008958:Models, Molecular; D008968:Molecular Conformation; D010957:Plasmids / Q000378:metabolism; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D006180:Proton-Translocating ATPases / Q000737:chemistry* / Q000378:metabolism; D013997:Time Factors
chemical_list: D000900:Anti-Bacterial Agents; D003432:Cross-Linking Reagents; D008024:Ligands; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases
references: Biochemistry. 1998 Nov 17;37(46):16423-9
references: 9819235
references: Eur J Biochem. 1992 Oct 15;209(2):681-7
references: 1425675
references: Biochim Biophys Acta. 2009 Aug;1787(8):1016-23
references: 19269272
references: Biochem Soc Trans. 1995 Nov;23(4):736-41
references: 8654828
references: Adv Exp Med Biol. 2012;726:5-16
references: 22297508
references: Anal Biochem. 1986 Nov 15;159(1):35-42
references: 2433963
references: Annu Rev Biochem. 1997;66:717-49
references: 9242922
references: J Biol Chem. 2007 Jul 13;282(28):20698-704
references: 17517893
references: Nature. 1970 Aug 15;227(5259):680-5
references: 5432063
references: J Biol Chem. 2008 Sep 12;283(37):25273-25280
references: 18625705
references: Proc Natl Acad Sci U S A. 2001 Nov 20;98(24):13649-54
references: 11707579
references: Structure. 2007 Aug;15(8):904-14
references: 17697996
references: Biochemistry. 2001 Sep 4;40(35):10664-70
references: 11524011
references: J Biol Chem. 2009 Jun 26;284(26):17457-64
references: 19411254
references: Proc Natl Acad Sci U S A. 2008 Nov 18;105(46):17760-5
references: 19001275
references: J Comput Chem. 2004 Oct;25(13):1605-12
references: 15264254
references: Biochemistry. 1987 Jul 14;26(14):4488-93
references: 2889464
references: Anal Biochem. 1977 Dec;83(2):346-56
references: 603028
references: Nat Struct Biol. 2000 Nov;7(11):1055-61
references: 11062563
references: Biochem J. 1982 Jan 15;202(1):15-23
references: 6211171
references: FEBS Lett. 2009 Apr 2;583(7):1121-6
references: 19265694
references: J Biol Chem. 2000 Sep 29;275(39):30157-62
references: 11001951
references: Nat Struct Mol Biol. 2004 Feb;11(2):142-8
references: 14730353
references: Biochim Biophys Acta. 1995 Oct 10;1231(3):275-81
references: 7578215
references: Cell Metab. 2008 Jul;8(1):13-25
references: 18590689
references: EMBO J. 2006 Nov 15;25(22):5433-42
references: 17082766
references: Proc Natl Acad Sci U S A. 2007 Jul 3;104(27):11233-8
references: 17581881
references: Gene. 1996 Feb 22;169(1):59-64
references: 8635750
references: Science. 2005 Jan 14;307(5707):223-7
references: 15591164
references: Structure. 1997 Sep 15;5(9):1219-30
references: 9331422
references: Proc Natl Acad Sci U S A. 2008 Dec 30;105(52):20722-7
references: 19075235
references: Proc Natl Acad Sci U S A. 1995 Nov 21;92(24):10964-8
references: 7479919
references: Proc Natl Acad Sci U S A. 2005 Mar 22;102(12):4288-93
references: 15758075
references: J Biol Chem. 2006 Jan 6;281(1):501-7
references: 16267041
references: Biochemistry. 1995 Dec 19;34(50):16412-8
references: 8845368
references: Cell. 2007 Jul 27;130(2):309-21
references: 17662945
references: Photosynth Res. 2004;79(3):319-29
references: 16328798
references: Biochemistry. 1991 Nov 12;30(45):10987-91
references: 1834172
references: J Biol Chem. 1960 Nov;235:3322-9
references: 13738472
references: Proc Natl Acad Sci U S A. 2008 Jan 29;105(4):1192-7
references: 18216260
references: J Biol Chem. 1999 Oct 1;274(40):28351-5
references: 10497194
references: J Biol Chem. 1998 Oct 9;273(41):26645-51
references: 9756905
references: Proc Natl Acad Sci U S A. 2006 Jun 6;103(23):8646-9
references: 16728506
references: J Biol Chem. 2010 Dec 31;285(53):42058-67
references: 20974856
references: Biol Chem Hoppe Seyler. 1994 Jan;375(1):43-51
references: 8003256
references: Biophys J. 2010 Apr 7;98(7):1227-36
references: 20371322
references: Protein Sci. 2008 Sep;17(9):1475-85
references: 18535304
references: Proc Natl Acad Sci U S A. 2007 Oct 2;104(40):15671-6
references: 17895376
references: Eur J Biochem. 1986 Dec 1;161(2):513-8
references: 2877881
references: J Biol Chem. 1992 Sep 15;267(26):18953-60
references: 1388160
references: J Biol Chem. 1990 Sep 25;265(27):16280-7
references: 2144528
references: Biophys J. 2008 Nov 15;95(10):4979-87
references: 18723591
references: Anal Biochem. 2008 Jun 15;377(2):209-17
references: 18405656
references: J Biol Chem. 2011 Apr 15;286(15):13423-9
references: 21345803
references: Biochemistry. 1991 Aug 27;30(34):8305-10
references: 1832012
references: Biochemistry. 1980 Feb 5;19(3):526-31
references: 6444514
references: Cell. 2001 Aug 10;106(3):331-41
references: 11509182
references: Biochemistry. 1991 Feb 5;30(5):1278-84
references: 1825019
references: Proc Natl Acad Sci U S A. 2010 Sep 28;107(39):16823-7
references: 20847295
references: N Engl J Med. 2009 Jun 4;360(23):2397-405
references: 19494215
references: J Biol Chem. 1982 Jul 10;257(13):7354-9
references: 6211441
references: J Biol Chem. 2007 Jan 5;282(1):764-72
references: 17092944
references: J Biol Chem. 2010 Apr 9;285(15):11411-7
references: 20154086
references: Biochim Biophys Acta. 1990 Oct 24;1020(1):43-8
references: 2145975
references: J Biol Chem. 1977 May 10;252(9):2891-9
references: 16006
references: J Biol Chem. 1999 Jul 2;274(27):19124-8
references: 10383416
references: FEMS Microbiol Lett. 2010 Dec;313(1):68-74
references: 21039782
references: EMBO J. 2006 Oct 4;25(19):4596-604
references: 16977308
references: Biochim Biophys Acta. 2006 May-Jun;1757(5-6):326-38
references: 16701076
references: EMBO J. 2005 Jun 15;24(12):2053-63
references: 15920483
references: J Biol Chem. 1993 Nov 5;268(31):23179-85
references: 8226836
references: J Biol Chem. 1994 Nov 18;269(46):28871-7
references: 7961847
references: Biochem Biophys Res Commun. 2006 Apr 14;342(3):800-7
references: 16517239
references: FEBS Lett. 1985 Mar 25;182(2):419-24
references: 2858407
references: J Biol Chem. 1993 Oct 5;268(28):20762-7
references: 8407901
references: Proc Natl Acad Sci U S A. 2012 Jul 10;109(28):11139-43
references: 22733764
references: Nat Struct Mol Biol. 2011 Jun;18(6):701-7
references: 21602818