A Phosphomimetic Mutation Stabilizes SOD1 and Rescues Cell Viability in the Context of an ALS-Associated Mutation.
The majority of amyotrophic lateral sclerosis (ALS)-related mutations in the enzyme Cu,Zn superoxide dismutase (SOD1), as well as a post-translational modification, glutathionylation, destabilize the protein and lead to a misfolded oligomer that is toxic to motor neurons. The biophysical role of another physiological SOD1 modification, T2-phosphorylation, has remained a mystery. Here, we find that a phosphomimetic mutation, T2D, thermodynamically stabilizes SOD1 even in the context of a strongly SOD1-destabilizing mutation, A4V, one of the most prevalent and aggressive ALS-associated mutations in North America. This stabilization protects against formation of toxic SOD oligomers and positively impacts motor neuron survival in cellular assays. We solve the crystal structure of T2D-SOD1 and explain its stabilization effect using discrete molecular dynamics (DMD) simulations. These findings imply that T2-phosphorylation may be a plausible innate cellular protection response against SOD1-induced cytotoxicity, and stabilizing the SOD1 native conformation might offer us viable pharmaceutical strategies against currently incurable ALS.
Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, NC 27599, USA; Program in Molecular and Cellular Biophysics, University of North Carolina, Chapel Hill, NC 27599, USA.
James M
JM
Fay
Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, NC 27599, USA.
Cheng
C
Zhu
Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, NC 27599, USA; Program in Molecular and Cellular Biophysics, University of North Carolina, Chapel Hill, NC 27599, USA; Curriculum in Bioinformatics and Computational Biology, University of North Carolina, Chapel Hill, NC 27599, USA.
Elizabeth A
EA
Proctor
Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, NC 27599, USA.
Yazhong
Y
Tao
Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, NC 27599, USA.
Wenjun
W
Cui
Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, NC 27599, USA.
Hengming
H
Ke
Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, NC 27599, USA; Program in Molecular and Cellular Biophysics, University of North Carolina, Chapel Hill, NC 27599, USA; Curriculum in Bioinformatics and Computational Biology, University of North Carolina, Chapel Hill, NC 27599, USA. Electronic address: dokh@unc.edu.
Nikolay V
NV
Dokholyan
D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics*; D002460:Cell Line; D002470:Cell Survival; D002942:Circular Dichroism; D004795:Enzyme Stability; D006801:Humans; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D009154:Mutation; D009474:Neurons / Q000166:cytology*; D010766:Phosphorylation; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D017433:Protein Structure, Secondary; D000072105:Superoxide Dismutase-1 / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism
C000606290:SOD1 protein, human; D000072105:Superoxide Dismutase-1
Neurobiol Dis. 2000 Dec;7(6 Pt B):623-43
11114261
Nat Rev Neurosci. 2001 Nov;2(11):806-19
11715057
Acta Neuropathol. 2002 Jan;103(1):26-35
11837744
J Neurosci Res. 2002 Sep 1;69(5):567-77
12210822
Brain. 2004 Jan;127(Pt 1):73-88
14534160
Dev Dyn. 1992 Jul;194(3):209-21
1467557
J Mol Graph Model. 2004 May;22(5):377-95
15099834
Proc Natl Acad Sci U S A. 2004 Oct 19;101(42):15094-9
15475574
Proc Natl Acad Sci U S A. 2006 Feb 28;103(9):3147-52
16488975
Biochim Biophys Acta. 2006 Nov-Dec;1762(11-12):1051-67
16713195
Nat Rev Neurosci. 2006 Sep;7(9):710-23
16924260
Neuron. 2006 Oct 5;52(1):39-59
17015226
Nat Rev Mol Cell Biol. 2007 Feb;8(2):101-12
17245412
Nat Methods. 2007 Jun;4(6):466-7
17538626
Proc Natl Acad Sci U S A. 2007 Aug 28;104(35):14157-62
17715066
Structure. 2007 Dec;15(12):1567-76
18073107
Mol Interv. 2007 Dec;7(6):313-24
18199853
Structure. 2008 Jul;16(7):1010-8
18611374
Proc Natl Acad Sci U S A. 2008 Dec 16;105(50):19696-701
19052230
J Biol Chem. 2009 May 15;284(20):13940-7
19299510
J Cell Biol. 2009 Dec 14;187(6):761-72
19951898
Free Radic Biol Med. 2010 Mar 1;48(5):629-41
19969067
Acta Crystallogr A. 1991 Mar 1;47 ( Pt 2):110-9
2025413
Nat Neurosci. 2010 Nov;13(11):1396-403
20953194
Proteins. 2011 Jan;79(1):261-70
21058396
J Mol Biol. 2011 May 6;408(3):555-67
21396374
Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):355-67
21460454
Biochemistry. 2011 Aug 16;50(32):7057-66
21739997
J Cell Biol. 2011 Aug 22;194(4):551-65
21859860
J Mol Biol. 2012 Aug 24;421(4-5):548-60
22210350
J Neurosci. 2012 Feb 1;32(5):1536-44
22302797
Prog Mol Biol Transl Sci. 2012;107:215-62
22482452
Hum Mutat. 2012 Sep;33(9):1345-51
22753137
Curr Top Med Chem. 2012;12(22):2504-22
23339304
Beilstein J Org Chem. 2013 Apr 15;9:717-32
23766784
Biophys J. 2013 Nov 5;105(9):2141-8
24209859
Sci Rep. 2013 Nov 20;3:3275
24253732
Proc Natl Acad Sci U S A. 2014 Jan 7;111(1):197-201
24344300
Nat Neurosci. 2014 Jan;17(1):17-23
24369373
Nat Commun. 2014 Mar 19;5:3446
24647101
Biochemistry. 2014 Apr 15;53(14):2423-32
24660965
J Biol Chem. 2014 Oct 10;289(41):28527-38
25164820
J Biol Chem. 2014 Oct 31;289(44):30690-701
25237191
Nat Commun. 2014 Nov 27;5:5502
25429517
Science. 2015 Mar 27;347(6229):1436-41
25700176
Neuron. 2015 Apr 8;86(1):218-32
25801706
Protein Sci. 2015 Dec;24(12):2081-9
26362407
J Biol Chem. 2015 Dec 18;290(51):30624-36
26511321
J Biol Chem. 2016 Feb 19;291(8):4144-55
26694608
Biochemistry. 2016 Jan 26;55(3):519-33
26710831
Proc Natl Acad Sci U S A. 2016 Jan 19;113(3):614-9
26719414
Nat Commun. 2016 Apr 13;7:11359
27072999
Methods Enzymol. 1997;276:307-26
27754618
Methods Enzymol. 1997;276:581-594
27799116
Biochim Biophys Acta. 1972 Dec 7;289(2):276-83
4346514
J Biol Chem. 1984 Apr 25;259(8):5301-5
6425287
Neurosci Lett. 1994 Sep 26;179(1-2):149-52
7845611
Science. 1993 Aug 20;261(5124):1047-51
8351519
Nature. 1993 Mar 4;362(6415):59-62
8446170
Dev Neurosci. 1996;18(5-6):492-8
8940623
J Histochem Cytochem. 1976 Jan;24(1):24-33
943439
Methods Cell Biol. 1998;57:265-78
9648110
J Neurochem. 1999 Feb;72(2):693-9
9930742