5 research outputs found
Dynamical Behavior of Human α-Synuclein Studied by Quasielastic Neutron Scattering
<div><p>α-synuclein (αSyn) is a protein consisting of 140 amino acid residues and is abundant in the presynaptic nerve terminals in the brain. Although its precise function is unknown, the filamentous aggregates (amyloid fibrils) of αSyn have been shown to be involved in the pathogenesis of Parkinson's disease, which is a progressive neurodegenerative disorder. To understand the pathogenesis mechanism of this disease, the mechanism of the amyloid fibril formation of αSyn must be elucidated. Purified αSyn from bacterial expression is monomeric but intrinsically disordered in solution and forms amyloid fibrils under various conditions. As a first step toward elucidating the mechanism of the fibril formation of αSyn, we investigated dynamical behavior of the purified αSyn in the monomeric state and the fibril state using quasielastic neutron scattering (QENS). We prepared the solution sample of 9.5 mg/ml purified αSyn, and that of 46 mg/ml αSyn in the fibril state, both at pD 7.4 in D<sub>2</sub>O. The QENS experiments on these samples were performed using the near-backscattering spectrometer, BL02 (<i>DNA</i>), at the Materials and Life Science Facility at the Japan Accelerator Research Complex, Japan. Analysis of the QENS spectra obtained shows that diffusive global motions are observed in the monomeric state but largely suppressed in the fibril state. However, the amplitude of the side chain motion is shown to be larger in the fibril state than in the monomeric state. This implies that significant solvent space exists within the fibrils, which is attributed to the αSyn molecules within the fibrils having a distribution of conformations. The larger amplitude of the side chain motion in the fibril state than in the monomeric state implies that the fibril state is entropically favorable.</p></div
Summary of parameters estimated from the fits to the EISF curves.
<p>The parameters calculated using <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0151447#pone.0151447.e002" target="_blank">Eq 2</a>, shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0151447#pone.0151447.g004" target="_blank">Fig 4</a> are shown. The parameters shown are (a) the fraction of frozen atoms (<i>p</i> in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0151447#pone.0151447.e002" target="_blank">Eq 2</a>), and (b) the radius of the confined sphere (<i>a</i> in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0151447#pone.0151447.e002" target="_blank">Eq 2</a>).</p
Examples of quasielastic neutron scattering spectra, S(Q,ω).
<p>The spectra of the samples, the solvent, and the difference spectra between the sample and the solvent, of αSyn in (a) the monomeric state and (b) the fibril state, at Q = 1.225 Å<sup>-1</sup> and at 280 K, are shown. Filled symbols in black and open symbols in blue show the spectra of the sample solutions and those of the solvent, respectively. The spectra of the empty cell were subtracted from these spectra. Filled symbols in red show the difference spectra between the sample and the solvent. (c) Fits using <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0151447#pone.0151447.e001" target="_blank">Eq 1</a> to the difference spectra, which are due to monomeric αSyn molecules in the sample solution. (d) Fits to the difference spectra, which are due to αSyn molecules in the fibril state. In the upper panels, open squares denote the data, thick solid lines in blue denote the total fits, solid lines in green and red denote the narrow and wide Lorentzian functions, corresponding to <i>L</i><sub>global</sub>(Q,ω) and <i>L</i><sub>local</sub>(Q,ω), respectively, thin solid lines in blue show the background, and dashed lines in black show the resolution functions. The lower panels show the residuals of the fits. (e) Reduced-χ<sup>2</sup> of the fits to the data. Filled squares and open squares are for the values to the data of αSyn in the monomeric state and the fibril state, respectively, at 280 K. Note that similar values of Reduced-χ<sup>2</sup> were obtained for the data measured at other temperatures.</p
The EISF curves of αSyn in (a) the monomeric state and (b) the fibril state.
<p>Solid lines are the fits with <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0151447#pone.0151447.e002" target="_blank">Eq 2</a>. The error bars are within in symbols where not shown.</p
Ligation-Dependent Picosecond Dynamics in Human Hemoglobin As Revealed by Quasielastic Neutron Scattering
Hemoglobin,
the vital O<sub>2</sub> carrier in red blood cells,
has long served as a classic example of an allosteric protein. Although
high-resolution X-ray structural models are currently available for
both the deoxy tense (T) and fully liganded relaxed (R) states of
hemoglobin, much less is known about their dynamics, especially on
the picosecond to subnanosecond time scales. Here, we investigate
the picosecond dynamics of the deoxy and CO forms of human hemoglobin
using quasielastic neutron scattering under near physiological conditions
in order to extract the dynamics changes upon ligation. From the analysis
of the global motions, we found that whereas the apparent diffusion
coefficients of the deoxy form can be described by assuming translational
and rotational diffusion of a rigid body, those of the CO form need
to involve an additional contribution of internal large-scale motions.
We also found that the local dynamics in the deoxy and CO forms are
very similar in amplitude but are slightly lower in frequency in the
former than in the latter. Our results reveal the presence of rapid
large-scale motions in hemoglobin and further demonstrate that this
internal mobility is governed allosterically by the ligation state
of the heme group