N-acetylation of α-synuclein enhances synaptic vesicle clustering mediated by α-synuclein and lysophosphatidylcholine
- Interdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, China
- University of Chinese Academy of Sciences, China
- Department of Molecular and Cellular Physiology, Stanford University, United States
- Department of Cancer Biology, University of Cincinnati College of Medicine, United States
- Bio-X Institutes, Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders (Ministry of Education), Shanghai Jiao Tong University, China
- Zhangjiang Institute for Advanced Study, Shanghai Jiao Tong University, China
- Brain and Mind Research Institute & Appel Institute for Alzheimer’s Disease Research, Weill Cornell Medicine, United States
- State Key Laboratory of Chemical Biology, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, China
Figures
Figure 1
N-terminal acetylation enhances synaptic vesicle (SV) clustering induced by α-synuclein (α-syn).
(a) α-syn is N-terminally acetylated in cells. Upper panel: Schematic representation of the delivery of 15N-labeled N-terminally unmodified α-syn (un-α-syn) into mammalian cells through …
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Figure 1—source data 1
The DLS numerical data.
- https://cdn.elifesciences.org/articles/97228/elife-97228-fig1-data1-v1.xlsx
Figure 2
Lysophosphatidylcholine (LPC) mediates the enhancement of vesicle clustering by N-terminal acetylation of α-synuclein (α-syn).
(a) Scheme of single-vesicle clustering assay for the functional study of Ac-α-syn on vesicle clustering. Vesicles were prepared with different amounts of LPC, and were labeled with DiD or remained …
Figure 3
N-terminal acetylation increases the α-synuclein (α-syn)–lysophosphatidylcholine (LPC) interaction.
Comparisons of residue-resolved nuclear magnetic resonance (NMR) signal intensity ratios (I/I0) of un-α-syn (upper) and Ac-α-syn (lower) during titration with LPC micelles (a), LPC-containing …
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Figure 3—source data 1
The NMR titration numerical data.
- https://cdn.elifesciences.org/articles/97228/elife-97228-fig3-data1-v1.xlsx
Figure 4
Ac-α-syn binding on lysophosphatidylcholine (LPC) shows high intermolecular interactions.
(a) The cross-linking patterns of Ac-α-syn in the presence of LPC and dioleoyl-phosphoserine (DOPS) mapped by mass spectrometry (MS) at the protein/lipid molar ratio of 1:50. Lines present the …
Author response image 1
Representative raw dataset of α-Syn-mediated synaptic vesicle (SV) clustering monitored by dynamic light scattering (DLS).
The gray-colored rows represent small particles (< 5 nm) that contributed zero to the particle number count.
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Escherichia coli) | BL21(DE3) | BioRad | 156–3003 | Electrocompetent cells |
| Cell line (Homo-sapiens) | HEK-293T epithelial-like cells | ATCC | CRL-3216 | Cell line authentication services (STR profiling) are offered by ATCC, not detected mycoplasma contamination |
| Transfected construct (human) | α-syn, Δ30 α-syn to pET22 vector | This paper; Zhao et al., 2024 | Constructs saved in C. Liu lab | |
| Commercial assay or kit | Neon transfection system kit | Invitrogen | MPK5000 | |
| Chemical compound, drug | 16:0 LPC, DOPC, DOPS, POPC, POPE, biotin-DPPE, cholesterol | Avanti Polar Lipids | 855675, 850375, 840035, 850457, 850757, 870277, 70000 | |
| Chemical compound, drug | Disuccinimidyl suberate (DSS) | Thermo Scientific | 21658 | |
| Chemical compound, drug | uranyl acetate | Sigma Aldrich | CDS021290 | |
| Chemical compound, drug | DiD | Invitrogen | D307 | |
| Software, algorithm | pLink | pLink | V1.9 | For XL-MS data |
| Software, algorithm | SPARKY | SPARKY | V3.115 | For NMR data |
| Software, algorithm | NMRpipe | NMRpipe | Build2018 | For NMR data |
| Software, algorithm | smCamera | TJ Ha’s lab | For single vesicle data | |
| Software, algorithm | Dynamics | Wyatt | V7.0 | For DLS data |
| Other | C57BL6 mice | Lingchang Shanghai | 8 wk old, male |
Additional files
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Supplementary file 1
Supplementary tables.
- https://cdn.elifesciences.org/articles/97228/elife-97228-supp1-v1.docx
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MDAR checklist
- https://cdn.elifesciences.org/articles/97228/elife-97228-mdarchecklist1-v1.docx
