α-Synuclein Misfolding: Single Molecule AFM Force Spectroscopy Study

Yu, Junping; Málková, Šárka; Lyubchenko, Yuri L.

doi:10.1016/j.jmb.2008.10.006

Cited by 74 publications

(139 citation statements)

References 54 publications

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“…Such stable a-Syn dimers were also identified in the AFM force spectroscopy experiments, but only at pH 2.7, whereas the fluorescence experiments used here were performed at neutral pH. The yield of rupture events of force measurements at neutral pH was very low, but measurements at pH 5.1 provided only one lifetime, t ¼ 0.23 s (19). The fluorescence data show that the population of type 2 conformers of dimers is 10%, so their detection with AFM force spectroscopy is problematic.…”

Section: Conformational Dynamics Of A-syn Dimersmentioning

confidence: 83%

“…Indeed, a similar immobilization of a-Syn was used in our prior AFM force spectroscopy studies, in which the interactions between two immobilized a-Syn proteins were probed (11,19,22,24). The same terminal cysteine was used for fluorescence labeling of a-Syn that was free in solution in the experimental setup (Fig.…”

Section: Biophysical Journal 108(8) 2038-2047mentioning

confidence: 99%

“…Using this approach, we demonstrated that the protein can form dimers with lifetimes in the range of a second that correspond to a barrier height as large as~28 k B T (Table S3). Recently, the AFM force spectroscopy approach was applied to characterize a-Syn dimers (11,19,22), and the value of the dimer lifetime was also on a timescale in the range of seconds. However, AFM force spectroscopy probes dimer stability along the direction of the applied force.…”

Section: Conformational Dynamics Of A-syn Dimersmentioning

confidence: 99%

“…Recently, our group developed an AFM-based SMFS approach designed to monitor the interaction between monomers tethered to the surface and the AFM tip (11,(19)(20)(21)(22)(23)(24)(25)(26)(27)(28). Using this approach, misfolded states of a-Syn and other proteins have been probed to reveal a number of common features.…”

Section: Introductionmentioning

confidence: 99%

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Direct Detection of α-Synuclein Dimerization Dynamics: Single-Molecule Fluorescence Analysis

Krasnoslobodtsev

Zhang

et al. 2015

Biophysical Journal

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The aggregation of a-synuclein (a-Syn) is linked to Parkinson's disease. The mechanism of early aggregation steps and the effect of pathogenic single-point mutations remain elusive. We report here a single-molecule fluorescence study of a-Syn dimerization and the effect of mutations. Specific interactions between tethered fluorophore-free a-Syn monomers on a substrate and fluorophore-labeled monomers diffusing freely in solution were observed using total internal reflection fluorescence microscopy. The results showed that wild-type (WT) a-Syn dimers adopt two types of dimers. The lifetimes of type 1 and type 2 dimers were determined to be 197 5 3 ms and 3334 5 145 ms, respectively. All three of the mutations used, A30P, E46K, and A53T, increased the lifetime of type 1 dimer and enhanced the relative population of type 2 dimer, with type 1 dimer constituting the major fraction. The kinetic stability of type 1 dimers (expressed in terms of lifetime) followed the order A30P (693 5 14 ms) > E46K (292 5 5 ms) > A53T (226 5 6 ms) > WT (197 5 3 ms). Type 2 dimers, which are more stable, had lifetimes in the range of several seconds. The strongest effect, observed for the A30P mutant, resulted in a lifetime 3.5 times higher than observed for the WT type 1 dimer. This mutation also doubled the relative fraction of type 2 dimer. These data show that single-point mutations promote dimerization, and they suggest that the structural heterogeneity of a-Syn dimers could lead to different aggregation pathways.

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Section: Conformational Dynamics Of A-syn Dimersmentioning

confidence: 83%

Section: Biophysical Journal 108(8) 2038-2047mentioning

confidence: 99%

Section: Conformational Dynamics Of A-syn Dimersmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Direct Detection of α-Synuclein Dimerization Dynamics: Single-Molecule Fluorescence Analysis

Krasnoslobodtsev

Zhang

et al. 2015

Biophysical Journal

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“…For example, covalent attachment of amyloid fibrils on the AFM tip for force spectroscopy, which will provide high accuracy and efficiency of binding, has rarely been used (Ganchev, et al, 2008;Yu et al, 2008). We expect more sophisticated AFM technique like tip modification, dynamic force spectroscopy, etc., would push AFM based research on amyloid mechanical properties to more fundamental level, single molecule force spectroscopy (SMFS), for example (Figure 11 (d)).…”

Section: Discussionmentioning

confidence: 99%

Nanomechanics of Amyloid Materials Studied by Atomic Force Microscopy

Zeng¹,

Duan²,

Besenbacher³

et al. 2012

Atomic Force Microscopy Investigations Into Biology - From Cell to Protein

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Alteration of the α‐Synuclein Folding Landscape by a Mutation Related to Parkinson’s Disease

et al. 2010

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Krankhafte Verformungen: Eine Einzelmolekül‐Fluoreszenztechnik wurde bei Strukturuntersuchungen an einem intrinsisch fehlgeordneten Gehirnprotein angewendet. Bei einer Mutation wurde festgestellt, dass sie die gekoppelte Bindungs‐ und Faltungsenergie des Proteins veränderte und das Schalten zwischen induzierten geordneten Strukturen verhinderte (siehe Bild). Die Beobachtungen liefern grundlegende Informationen zu molekularen Vorgängen bei der Parkinson‐Krankheit.

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α-Synuclein Misfolding: Single Molecule AFM Force Spectroscopy Study

Cited by 74 publications

References 54 publications

Direct Detection of α-Synuclein Dimerization Dynamics: Single-Molecule Fluorescence Analysis

Direct Detection of α-Synuclein Dimerization Dynamics: Single-Molecule Fluorescence Analysis

Nanomechanics of Amyloid Materials Studied by Atomic Force Microscopy

Alteration of the α‐Synuclein Folding Landscape by a Mutation Related to Parkinson’s Disease

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