Noemi Ferrante Carrante
Doctoral student
Transient Interactions of α‑Synuclein N- and C‑Termini
Author
Summary, in English
α-Synuclein (αSyn) is a neuronal protein predominantly found at the
synapse, involved in vesicle trafficking. αSyn aggregates are also the
main component of Lewy bodies, the hallmarks of Parkinson’s disease.
Interactions between the N- and C-termini of αSyn play crucial roles in
its behavior in solution, membrane binding, and aggregation. Studying
these interactions provides valuable insights into the physiological and
pathological functions of αSyn. Here, we employed photoinduced
cross-linking of unmodified proteins (PICUP) to identify the transient
contacts of αSyn in different conformational states. By using
tyrosine-to-phenylalanine mutations to block the reactivity of specific
amino acids, we establish key cross-links in each state. In solution, we
identify internal contacts between the N- and C-termini of monomers, as
well as intermonomer contacts between C-termini in oligomers. When αSyn
is bound to membranes, the internal cross-linking is blocked, while the
cross-linking between C-terminal regions persists. In fibrils,
cross-linking is significantly reduced, primarily occurring between the
C-termini of adjacent monomers. This work highlights the effectiveness
of PICUP for reporting on the transient contacts involved in αSyn
self-assembly and its coassembly with lipid membranes, while providing a
streamlined protocol that opens avenues for studying protein–protein
interactions for a wide range of systems.
synapse, involved in vesicle trafficking. αSyn aggregates are also the
main component of Lewy bodies, the hallmarks of Parkinson’s disease.
Interactions between the N- and C-termini of αSyn play crucial roles in
its behavior in solution, membrane binding, and aggregation. Studying
these interactions provides valuable insights into the physiological and
pathological functions of αSyn. Here, we employed photoinduced
cross-linking of unmodified proteins (PICUP) to identify the transient
contacts of αSyn in different conformational states. By using
tyrosine-to-phenylalanine mutations to block the reactivity of specific
amino acids, we establish key cross-links in each state. In solution, we
identify internal contacts between the N- and C-termini of monomers, as
well as intermonomer contacts between C-termini in oligomers. When αSyn
is bound to membranes, the internal cross-linking is blocked, while the
cross-linking between C-terminal regions persists. In fibrils,
cross-linking is significantly reduced, primarily occurring between the
C-termini of adjacent monomers. This work highlights the effectiveness
of PICUP for reporting on the transient contacts involved in αSyn
self-assembly and its coassembly with lipid membranes, while providing a
streamlined protocol that opens avenues for studying protein–protein
interactions for a wide range of systems.
Department/s
- Biochemistry and Structural Biology
- MultiPark: Multidisciplinary research on neurodegenerative diseases
- LU Profile Area: Light and Materials
- LTH Profile Area: Nanoscience and Semiconductor Technology
- NanoLund: Centre for Nanoscience
- Physical Chemistry
- LU Profile Area: Proactive Ageing
Publishing year
2026-03-30
Language
English
Publication/Series
ACS Chemical Neuroscience
Document type
Journal article
Publisher
The American Chemical Society (ACS)
Topic
- Biophysics
Status
Epub
ISBN/ISSN/Other
- ISSN: 1948-7193