title: Familial Parkinson disease-associated mutations alter the site-specific microenvironment and dynamics of α-synuclein.
abstract: Human α-synuclein (α-Syn) is a natively unstructured protein whose aggregation into amyloid fibrils is associated with Parkinson disease (PD) pathogenesis. Mutations of α-Syn, E46K, A53T, and A30P, have been linked to the familial form of PD. In vitro aggregation studies suggest that increased propensity to form non-fibrillar oligomers is the shared property of these familial PD-associated mutants. However, the structural basis of the altered aggregation propensities of these PD-associated mutants is not yet clear. To understand this, we studied the site-specific structural dynamics of wild type (WT) α-Syn and its three PD mutants (A53T, E46K, and A30P). Tryptophan (Trp) was substituted at the N terminus, central hydrophobic region, and C terminus of all α-Syns. Using various biophysical techniques including time-resolved fluorescence studies, we show that irrespective of similar secondary structure and early oligomerization propensities, familial PD-associated mutations alter the site-specific microenvironment, solvent exposure, and conformational flexibility of the protein. Our results further show that the common structural feature of the three PD-associated mutants is more compact and rigid sites at their N and C termini compared with WT α-Syn that may facilitate the formation of a partially folded intermediate that eventually leads to their increased oligomerization propensities.
authors: From the Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Mumbai 400 076, India and.
authors: Shruti
authors: S
authors: Sahay
authors: From the Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Mumbai 400 076, India and.
authors: Dhiman
authors: D
authors: Ghosh
authors: From the Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Mumbai 400 076, India and.
authors: Saumya
authors: S
authors: Dwivedi
authors: From the Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Mumbai 400 076, India and.
authors: Arunagiri
authors: A
authors: Anoop
authors: From the Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Mumbai 400 076, India and.
authors: Ganesh Maruti
authors: GM
authors: Mohite
authors: Department of Chemical Sciences, Tata Institute of Fundamental Research, Mumbai 400 005, India.
authors: Mamata
authors: M
authors: Kombrabail
authors: Department of Chemical Sciences, Tata Institute of Fundamental Research, Mumbai 400 005, India gk@tifr.res.in.
authors: Guruswamy
authors: G
authors: Krishnamoorthy
authors: From the Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Mumbai 400 076, India and samirmaji@iitb.ac.in.
authors: Samir K
authors: SK
authors: Maji
mesh_terms: D000595:Amino Acid Sequence; D006801:Humans; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010300:Parkinson Disease / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D051844:alpha-Synuclein / Q000737:chemistry / Q000378:metabolism*
chemical_list: D051844:alpha-Synuclein
keywords: Amyloid; Fluorescence Anisotropy; Parkinson Disease; Protein Aggregation; Protein Misfolding; α-Synuclein
references: Biochemistry. 2006 Feb 28;45(8):2752-60
references: 16489768
references: Biochemistry. 2014 Feb 11;53(5):807-9
references: 24450731
references: Neurobiol Aging. 2014 Sep;35(9):2180.e1-5
references: 24746362
references: J Biol Chem. 2014 Jun 13;289(24):16884-903
references: 24782311
references: Neurotoxicology. 2002 Oct;23(4-5):553-67
references: 12428728
references: Proc Natl Acad Sci U S A. 2003 Jan 7;100(1):330-5
references: 12506200
references: Neuron. 2000 Jan;25(1):239-52
references: 10707987
references: Biochemistry. 2000 Mar 14;39(10):2552-63
references: 10704204
references: Biophys J. 2012 Jun 20;102(12):2894-905
references: 22735540
references: Neurobiol Aging. 2012 Sep;33(9):2172-85
references: 22118903
references: Biochemistry. 2012 Nov 20;51(46):9260-9
references: 23110669
references: PLoS One. 2013;8(1):e53487
references: 23349712
references: ACS Chem Neurosci. 2013 Mar 20;4(3):486-97
references: 23374072
references: ACS Chem Neurosci. 2013 Mar 20;4(3):498-508
references: 23374074
references: ACS Chem Neurosci. 2013 Mar 20;4(3):393-407
references: 23509976
references: ACS Chem Neurosci. 2013 Jul 17;4(7):1101-13
references: 23607785
references: Parkinsonism Relat Disord. 2014 Jan;20 Suppl 1:S29-34
references: 24262183
references: Biochemistry. 2013 Oct 8;52(40):6925-7
references: 24047453
references: J Biol Chem. 2000 Jul 28;275(30):22645-9
references: 10806193
references: J Biol Chem. 2001 Apr 6;276(14):10737-44
references: 11152691
references: J Biol Chem. 2001 Jan 26;276(4):2380-6
references: 11060312
references: Nat Struct Biol. 2001 Sep;8(9):799-804
references: 11524685
references: J Biol Chem. 2001 Sep 14;276(37):35176-84
references: 11441003
references: Biochemistry. 2001 Sep 25;40(38):11604-13
references: 11560511
references: J Biol Chem. 2002 Apr 5;277(14):11970-8
references: 11812782
references: J Neuropsychiatry Clin Neurosci. 2002 Spring;14(2):223-36; discussion 222
references: 11983801
references: Nat Med. 2002 Jun;8(6):600-6
references: 12042811
references: Curr Opin Genet Dev. 2002 Jun;12(3):299-306
references: 12076673
references: J Neurosci. 2002 Oct 15;22(20):8797-807
references: 12388586
references: FEBS Lett. 1994 May 23;345(1):27-32
references: 8194594
references: Neuron. 1995 Feb;14(2):467-75
references: 7857654
references: Nature. 1997 Aug 28;388(6645):839-40
references: 9278044
references: Am J Pathol. 1998 Apr;152(4):879-84
references: 9546347
references: J Biol Chem. 1998 Apr 17;273(16):9443-9
references: 9545270
references: Biochemistry. 2003 Jan 28;42(3):672-8
references: 12534279
references: J Mol Biol. 2003 Jun 13;329(4):763-78
references: 12787676
references: Biochemistry. 2003 Jul 22;42(28):8530-40
references: 12859200
references: J Biomol Struct Dyn. 2003 Oct;21(2):211-34
references: 12956606
references: FASEB J. 2004 Apr;18(6):617-26
references: 15054084
references: FEBS Lett. 2004 Oct 22;576(3):363-8
references: 15498564
references: Annu Rev Biochem. 1985;54:43-71
references: 3896124
references: Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11282-6
references: 8248242
references: Proc Natl Acad Sci U S A. 2000 Jan 18;97(2):571-6
references: 10639120
references: Nature. 2000 Mar 23;404(6776):394-8
references: 10746727
references: Brain Res. 2000 Jun 2;866(1-2):33-43
references: 10825478
references: Chem Rev. 2006 May;106(5):1769-84
references: 16683754
references: Protein Pept Lett. 2006;13(4):385-90
references: 16712515
references: Acc Chem Res. 2006 Sep;39(9):628-34
references: 16981679
references: J Mol Biol. 2007 Mar 9;366(5):1510-22
references: 17222866
references: Biochemistry. 2007 Jun 19;46(24):7107-18
references: 17530780
references: Proc Natl Acad Sci U S A. 2008 Jun 24;105(25):8637-42
references: 18550842
references: Brain Res. 2008 Dec 9;1244:40-52
references: 18817762
references: Acta Neurobiol Exp (Wars). 2008;68(4):509-15
references: 19112474
references: Chembiochem. 2009 Jan 5;10(1):176-83
references: 19067456
references: J Mol Biol. 2009 May 22;388(5):1022-32
references: 19345692
references: J Mol Biol. 2009 Oct 30;393(3):735-52
references: 19716830
references: EMBO J. 2009 Oct 21;28(20):3256-68
references: 19745811
references: J Mol Biol. 2009 Dec 18;394(5):826-33
references: 19837084
references: Proc Natl Acad Sci U S A. 2009 Nov 24;106(47):20051-6
references: 19892735
references: Proc Natl Acad Sci U S A. 2011 Mar 8;108(10):4194-9
references: 21325059
references: Neuron. 2011 Oct 6;72(1):57-71
references: 21982369
references: Proc Natl Acad Sci U S A. 1998 May 26;95(11):6469-73
references: 9600990
references: Trends Neurosci. 1998 Jun;21(6):249-54
references: 9641537
references: Biochemistry. 1998 Jul 14;37(28):9976-82
references: 9665702
references: Nat Med. 1998 Nov;4(11):1318-20
references: 9809558
references: J Biol Chem. 1999 Jul 9;274(28):19509-12
references: 10391881
references: Proc Natl Acad Sci U S A. 2004 Nov 23;101(47):16466-71
references: 15536128
references: J Am Chem Soc. 2005 Jan 19;127(2):476-7
references: 15643843
references: Proc Natl Acad Sci U S A. 2005 Feb 1;102(5):1430-5
references: 15671169
references: J Biol Chem. 2005 Mar 4;280(9):7800-7
references: 15632170
references: J Biol Chem. 2005 Sep 2;280(35):30649-52
references: 16020550
references: Biochemistry. 2005 Oct 11;44(40):13365-76
references: 16201761