Structural Basis for Gating and Activation of RyR1.
The type-1 ryanodine receptor (RyR1) is an intracellular calcium (Ca(2+)) release channel required for skeletal muscle contraction. Here, we present cryo-EM reconstructions of RyR1 in multiple functional states revealing the structural basis of channel gating and ligand-dependent activation. Binding sites for the channel activators Ca(2+), ATP, and caffeine were identified at interdomain interfaces of the C-terminal domain. Either ATP or Ca(2+) alone induces conformational changes in the cytoplasmic assembly ("priming"), without pore dilation. In contrast, in the presence of all three activating ligands, high-resolution reconstructions of open and closed states of RyR1 were obtained from the same sample, enabling analyses of conformational changes associated with gating. Gating involves global conformational changes in the cytosolic assembly accompanied by local changes in the transmembrane domain, which include bending of the S6 transmembrane segment and consequent pore dilation, displacement, and deformation of the S4-S5 linker and conformational changes in the pseudo-voltage-sensor domain.
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA; Howard Hughes Medical Institute, Columbia University, New York, NY 10032, USA.
Amédée
A
des Georges
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA.
Oliver B
OB
Clarke
Department of Physiology and Cellular Biophysics, Columbia University, New York, NY 10032, USA.
Ran
R
Zalk
Department of Physiology and Cellular Biophysics, Columbia University, New York, NY 10032, USA.
Qi
Q
Yuan
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA; Howard Hughes Medical Institute, Columbia University, New York, NY 10032, USA.
Kendall J
KJ
Condon
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA; Howard Hughes Medical Institute, Columbia University, New York, NY 10032, USA.
Robert A
RA
Grassucci
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA; Department of Physiology and Cellular Biophysics, Columbia University, New York, NY 10032, USA. Electronic address: wayne@xtl.cumc.columbia.edu.
Wayne A
WA
Hendrickson
Department of Physiology and Cellular Biophysics, Columbia University, New York, NY 10032, USA; Department of Medicine, Columbia University, New York, NY 10032, USA; Wu Center for Molecular Cardiology, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA. Electronic address: arm42@cumc.columbia.edu.
Andrew R
AR
Marks
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA; Howard Hughes Medical Institute, Columbia University, New York, NY 10032, USA; Department of Biological Sciences, Columbia University, New York, NY 10032, USA. Electronic address: jf2192@cumc.columbia.edu.
Joachim
J
Frank
D000818:Animals; D001665:Binding Sites; D002110:Caffeine / Q000737:chemistry; D002118:Calcium / Q000737:chemistry; D002120:Calcium Channel Agonists / Q000737:chemistry*; D020285:Cryoelectron Microscopy; D015640:Ion Channel Gating; D008024:Ligands; D009119:Muscle Contraction; D000072417:Protein Domains; D011817:Rabbits; D019837:Ryanodine Receptor Calcium Release Channel / Q000737:chemistry*; D022021:Tacrolimus Binding Proteins / Q000737:chemistry
D002120:Calcium Channel Agonists; D008024:Ligands; D019837:Ryanodine Receptor Calcium Release Channel; D002110:Caffeine; D022021:Tacrolimus Binding Proteins; D002118:Calcium
RyR; calcium release; cryo-EM; excitation contraction coupling; gating; ryanodine; single-particle reconstruction; structure
Br J Pharmacol. 2000 Aug;130(7):1618-26
10928966
J Struct Biol. 2015 Nov;192(2):216-21
26278980
J Biol Chem. 1987 Dec 5;262(34):16636-43
2445748
J Mol Biol. 1988 Jan 5;199(1):137-47
2451023
Nature. 2015 Jan 1;517(7532):50-5
25517095
Bioinformatics. 2009 May 1;25(9):1189-91
19151095
Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):486-501
20383002
J Mol Graph. 1996 Dec;14(6):354-60, 376
9195488
J Mol Biol. 2003 Oct 31;333(4):721-45
14568533
Nat Commun. 2013;4:1506
23422674
J Biol Chem. 2000 Apr 21;275(16):11618-25
10766778
PLoS Biol. 2009 Apr 14;7(4):e85
19402748
J Biol Chem. 2004 Dec 17;279(51):53028-35
15469935
J Struct Biol. 2012 Dec;180(3):519-30
23000701
J Biol Chem. 1994 May 6;269(18):13305-12
8175760
Antioxid Redox Signal. 2000 Spring;2(1):41-5
11232598
Arch Biochem Biophys. 1991 Oct;290(1):239-47
1898095
Proc Natl Acad Sci U S A. 2002 Dec 24;99(26):16725-30
12486242
Biophys J. 1998 Mar;74(3):1296-304
9512027
J Gen Physiol. 2008 Dec;132(6):667-80
19029374
Biophys J. 1999 Sep;77(3):1528-39
10465763
Nat Methods. 2013 Jun;10(6):584-90
23644547
J Biol Chem. 1994 May 6;269(18):13076-9
8175731
J Physiol. 2001 Dec 15;537(Pt 3):763-78
11744753
Elife. 2015 May 29;4:e06980
26023829
Science. 2014 Dec 12;346(6215):1377-80
25504723
Nat Methods. 2014 Jan;11(1):63-5
24213166
J Biol Chem. 2016 Jan 29;291(5):2150-60
26663082
PLoS One. 2012;7(11):e48725
23144945
Nature. 2015 Jan 1;517(7532):39-43
25470059
J Comput Chem. 2004 Oct;25(13):1605-12
15264254
Mol Pharmacol. 2006 Jun;69(6):1990-7
16540598
Biochemistry. 1997 Mar 11;36(10):2939-50
9062124
J Biol Chem. 1994 Jun 3;269(22):15876-84
8195243
Nat Commun. 2014 Nov 05;5:5397
25370123
Protein J. 2009 Jun;28(5):240-6
19636685
Nucleic Acids Res. 2004 Mar 19;32(5):1792-7
15034147
Cell. 1994 May 20;77(4):513-23
7514503
Nature. 2015 Nov 19;527(7578):336-41
26458101
Mol Membr Biol. 2007 May-Jun;24(3):185-93
17520475
J Struct Biol. 2005 Jul;151(1):41-60
15890530
J Biol Chem. 2011 Oct 14;286(41):35571-7
21862589
Proc Natl Acad Sci U S A. 2001 Feb 27;98(5):2865-70
11226332
Biotechniques. 1992 Jun;12(6):870-4, 876-9
1642895
Cell Calcium. 1988 Feb;9(1):1-7
2452017
Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7521-5
1502163
J Struct Biol. 2015 Feb;189(2):114-22
25486611
Nat Commun. 2015 Aug 06;6:7947
26245150
Biophys J. 1999 Oct;77(4):1936-44
10512814
Nature. 2015 Jan 1;517(7532):44-9
25470061
PLoS One. 2013 Jun 12;8(6):e66542
23776685
Nature. 2010 Nov 25;468(7323):585-8
21048710
Science. 1998 Aug 7;281(5378):818-21
9694652
FEBS J. 2012 Oct;279(20):3952-64
22913516
Nature. 1991 Jun 27;351(6329):751-4
1648178
Circ Res. 1996 Jun;78(6):990-7
8635249
Mol Pharmacol. 1990 May;37(5):735-41
1692609
Mol Pharmacol. 1992 Dec;42(6):1049-57
1480132
Ultramicroscopy. 2013 Dec;135:24-35
23872039
J Gen Physiol. 2001 Jul;118(1):33-44
11429443
J Biol Chem. 2015 Jul 10;290(28):17535-45
25998124
Biophys J. 2006 Jan 1;90(1):173-82
16227507
Q Rev Biophys. 1988 May;21(2):129-228
3043536