Format

Send to

Choose Destination
See comment in PubMed Commons below
J Biol Chem. 2014 Apr 11;289(15):10727-37. doi: 10.1074/jbc.M113.524520. Epub 2014 Feb 24.

Molecular basis for preventing α-synuclein aggregation by a molecular tweezer.

Author information

1
From the Department of Physics and Astronomy, Michigan State University, East Lansing, Michigan 48823.

Abstract

Recent work on α-synuclein has shown that aggregation is controlled kinetically by the rate of reconfiguration of the unstructured chain, such that the faster the reconfiguration, the slower the aggregation. In this work we investigate this relationship by examining α-synuclein in the presence of a small molecular tweezer, CLR01, which binds selectively to Lys side chains. We find strong binding to multiple Lys within the chain as measured by fluorescence and mass-spectrometry and a linear increase in the reconfiguration rate with concentration of the inhibitor. Top-down mass-spectrometric analysis shows that the main binding of CLR01 to α-synuclein occurs at the N-terminal Lys-10/Lys-12. Photo-induced cross-linking of unmodified proteins (PICUP) analysis shows that under the conditions used for the fluorescence analysis, α-synuclein is predominantly monomeric. The results can be successfully modeled using a kinetic scheme in which two aggregation-prone monomers can form an encounter complex that leads to further oligomerization but can also dissociate back to monomers if the reconfiguration rate is sufficiently high. Taken together, the data provide important insights into the preferred binding site of CLR01 on α-synuclein and the mechanism by which the molecular tweezer prevents self-assembly into neurotoxic aggregates by α-synuclein and presumably other amyloidogenic proteins.

KEYWORDS:

Inhibitor; Intramolecular Diffusion; Mass Spectrometry (MS); Parkinson's Disease; Protein Aggregation; Spectroscopy; α-Synuclein

PMID:
24567327
PMCID:
PMC4036189
DOI:
10.1074/jbc.M113.524520
[Indexed for MEDLINE]
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for HighWire Icon for PubMed Central
    Loading ...
    Support Center