Structural characterization of the interaction of α-synuclein nascent chains with the ribosomal surface and trigger factor.
The ribosome is increasingly becoming recognized as a key hub for integrating quality control processes associated with protein biosynthesis and cotranslational folding (CTF). The molecular mechanisms by which these processes take place, however, remain largely unknown, in particular in the case of intrinsically disordered proteins (IDPs). To address this question, we studied at a residue-specific level the structure and dynamics of ribosome-nascent chain complexes (RNCs) of α-synuclein (αSyn), an IDP associated with Parkinson's disease (PD). Using solution-state nuclear magnetic resonance (NMR) spectroscopy and coarse-grained molecular dynamics (MD) simulations, we find that, although the nascent chain (NC) has a highly disordered conformation, its N-terminal region shows resonance broadening consistent with interactions involving specific regions of the ribosome surface. We also investigated the effects of the ribosome-associated molecular chaperone trigger factor (TF) on αSyn structure and dynamics using resonance broadening to define a footprint of the TF-RNC interactions. We have used these data to construct structural models that suggest specific ways by which emerging NCs can interact with the biosynthesis and quality control machinery.
Institute of Structural and Molecular Biology, University College London, London WC1E 6BT, United Kingdom; Institute of Structural and Molecular Biology, Birkbeck College, University of London, London WC1E 7HX, United Kingdom;
Annika
A
Deckert
Institute of Structural and Molecular Biology, University College London, London WC1E 6BT, United Kingdom; Institute of Structural and Molecular Biology, Birkbeck College, University of London, London WC1E 7HX, United Kingdom;
Christopher A
CA
Waudby
Institute of Structural and Molecular Biology, University College London, London WC1E 6BT, United Kingdom; Institute of Structural and Molecular Biology, Birkbeck College, University of London, London WC1E 7HX, United Kingdom;
Tomasz
T
Wlodarski
Institute of Structural and Molecular Biology, University College London, London WC1E 6BT, United Kingdom; Institute of Structural and Molecular Biology, Birkbeck College, University of London, London WC1E 7HX, United Kingdom;
Anne S
AS
Wentink
Institute of Structural and Molecular Biology, University College London, London WC1E 6BT, United Kingdom; Institute of Structural and Molecular Biology, Birkbeck College, University of London, London WC1E 7HX, United Kingdom;
Xiaolin
X
Wang
Institute of Structural and Molecular Biology, University College London, London WC1E 6BT, United Kingdom; Institute of Structural and Molecular Biology, Birkbeck College, University of London, London WC1E 7HX, United Kingdom;
John P
JP
Kirkpatrick
Institute of Structural and Molecular Biology, University College London, London WC1E 6BT, United Kingdom; Institute of Structural and Molecular Biology, Birkbeck College, University of London, London WC1E 7HX, United Kingdom;
Jack F S
JF
Paton
Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, United Kingdom.
Carlo
0000-0002-9923-8590
C
Camilloni
Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, United Kingdom.
Predrag
P
Kukic
Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, United Kingdom.
Christopher M
CM
Dobson
Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, United Kingdom.
Michele
M
Vendruscolo
Institute of Structural and Molecular Biology, University College London, London WC1E 6BT, United Kingdom; Institute of Structural and Molecular Biology, Birkbeck College, University of London, London WC1E 7HX, United Kingdom; l.cabrita@ucl.ac.uk j.christodoulou@ucl.ac.uk.
Lisa D
LD
Cabrita
Institute of Structural and Molecular Biology, University College London, London WC1E 6BT, United Kingdom; Institute of Structural and Molecular Biology, Birkbeck College, University of London, London WC1E 7HX, United Kingdom; l.cabrita@ucl.ac.uk j.christodoulou@ucl.ac.uk.
John
J
Christodoulou
D001665:Binding Sites; D003198:Computer Simulation; D008956:Models, Chemical; D062105:Molecular Docking Simulation; D011485:Protein Binding; D011487:Protein Conformation; D000072417:Protein Domains; D012270:Ribosomes / Q000737:chemistry* / Q000648:ultrastructure*; D013499:Surface Properties; D051844:alpha-Synuclein / Q000737:chemistry* / Q000648:ultrastructure*
D051844:alpha-Synuclein
NMR spectroscopy; cotranslational folding; nascent chain; ribosome; α-synuclein