Multiple Tight Phospholipid-Binding Modes of α-Synuclein Revealed by Solution NMR Spectroscopy

被引:310
作者
Bodner, Christina R. [1 ,2 ]
Dobson, Christopher M. [2 ]
Bax, Ad [1 ]
机构
[1] NIDDK, Chem Phys Lab, NIH, Bethesda, MD 20892 USA
[2] Univ Cambridge, Dept Chem, Cambridge CB2 1EW, England
基金
英国惠康基金;
关键词
alpha-helix; membrane binding; NOE; Parkinson's disease; SUV; DISEASE-LINKED MUTATIONS; A-BETA COMPONENT; NUCLEAR-MAGNETIC-RESONANCE; CENTRAL-NERVOUS-SYSTEM; PARKINSONS-DISEASE; ALZHEIMERS-DISEASE; LEWY BODIES; VESICLE PERMEABILIZATION; TRANSLATIONAL DIFFUSION; MEMBRANE INTERACTIONS;
D O I
10.1016/j.jmb.2009.05.066
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
'In dopaminergic neurons, alpha-synuclein (alpha S) partitions between a disordered cytosolic state and a lipid-bound state. Binding of alpha S to membrane phospholipids is implicated in its functional role in synaptic regulation, but also impacts fibril formation associated with Parkinson's disease. We describe here a solution NMR study in which alpha S is added to small unilamellar vesicles of a composition mimicking synaptic vesicles; the results provide evidence for multiple distinct phospholipid-binding modes of US. Exchange between the free state and the lipid-bound alpha S state, and between different bound states is slow on the NMR timescale, being in the range of 1-10 s(-1). Partitioning of the binding modes is dependent on lipid/alpha S stoichiometry, and tight binding with slow-exchange kinetics is observed at stoichiometries as low as 2:1. In all lipid-bound states, a segment of residues starting at the N-terminus of alpha S adopts an alpha-helical conformation, while succeeding residues retain the characteristics of a random coil. The 40 C-terminal residues remain dynamically disordered, even at high-lipid concentrations, but can also bind to lipids to an extent that appears to be determined by the fraction of cis X-Pro peptide bonds in this region. While lipid-bound alpha S exhibits dynamic properties that preclude its direct observation by NMR, its exchange with the NMR-visible free form allows for its indirect characterization. Rapid amide-amide nuclear Overhauser enhancement buildup points to a large et-helical conformation, and a distinct increase in fluorescence anisotropy attributed to Tyr39 indicates an ordered environment for this "dark state." Titration of alpha S with increasing amounts of lipids suggests that the binding mode under high-lipid conditions remains qualitatively similar to that in the low-lipid case. The NMR data appear incompatible with the commonly assumed model where alpha S lies in an alpha-helical conformation on the membrane surface and instead suggest that considerable remodeling of the vesicles is induced by alpha S. Published by Elsevier Ltd.
引用
收藏
页码:775 / 790
页数:16
相关论文
共 92 条
[81]   Comparison of structure and dynamics of micelle-bound human α-synuclein and Parkinson disease variants [J].
Ulmer, TS ;
Bax, A .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2005, 280 (52) :43179-43187
[82]   Structure and dynamics of micelle-bound human α-synuclein [J].
Ulmer, TS ;
Bax, A ;
Cole, NB ;
Nussbaum, RL .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2005, 280 (10) :9595-9603
[83]   Vesicle permeabilization by protofibrillar α-synuclein is sensitive to Parkinson's disease-linked mutations and occurs by a pore-like mechanism [J].
Volles, MJ ;
Lansbury, PT .
BIOCHEMISTRY, 2002, 41 (14) :4595-4602
[84]   Vesicle permeabilization by protofibrillar α-synuclein:: Implications for the pathogenesis and treatment of Parkinson's disease [J].
Volles, MJ ;
Lee, SJ ;
Rochet, JC ;
Shtilerman, MD ;
Ding, TT ;
Kessler, JC ;
Lansbury, PT .
BIOCHEMISTRY, 2001, 40 (26) :7812-7819
[85]   Mapping chemical exchange in proteins with MW > 50 kD [J].
Wang, CY ;
Rance, M ;
Palmer, AG .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2003, 125 (30) :8968-8969
[86]   NACP, a protein implicated in Alzheimer's disease and learning, is natively unfolded [J].
Weinreb, PH ;
Zhen, WG ;
Poon, AW ;
Conway, KA ;
Lansbury, PT .
BIOCHEMISTRY, 1996, 35 (43) :13709-13715
[87]   α-synuclein oligomerization:: a role for lipids? [J].
Welch, K ;
Yuan, JY .
TRENDS IN NEUROSCIENCES, 2003, 26 (10) :517-519
[88]   Delayed localization of synelfin (synuclein, NACP) to presynaptic terminals in cultured rat hippocampal neurons [J].
Withers, GS ;
George, JM ;
Banker, GA ;
Clayton, DF .
DEVELOPMENTAL BRAIN RESEARCH, 1997, 99 (01) :87-94
[89]   Helical α-synuclein forms highly conductive ion channels [J].
Zakharov, Stanislav D. ;
Hulleman, John D. ;
Dutseva, Elena A. ;
Antonenko, Yuri N. ;
Rochet, Jean-Christophe ;
Cramer, William A. .
BIOCHEMISTRY, 2007, 46 (50) :14369-14379
[90]   The new mutation, E46K, of α-synuclein causes Parkinson and Lewy body dementia [J].
Zarranz, JJ ;
Alegre, J ;
Gómez-Esteban, JC ;
Lezcano, E ;
Ros, R ;
Ampuero, I ;
Vidal, L ;
Hoenicka, J ;
Rodriguez, O ;
Atarés, B ;
Llorens, V ;
Tortosa, EG ;
del Ser, T ;
Muñoz, DG ;
de Yebenes, JG .
ANNALS OF NEUROLOGY, 2004, 55 (02) :164-173