Sources of aSyn detailed table (Recombinant / cell culture / Brain extract / Chemical synthetic with Method / pros / Product / parameter modelling rows), Skin aSyn (Syn-One test, Vasiliev 2018, Donadio 2024 #2752), Species / Native conformation, Primary aSyn species table (monomeric / tetramer / pSer129 / Misfolded / β-pleated sheet / Insoluble / Oligomeric / protofibril / amyloid-like fibril / Lewy bodies)
Sources of aSyn — detail table
(used review: Vandhamtoyu, 2022 (A.F.))
| Recombinant (bacterial expression) FL, PFF | cell culture | Brain extract | Chemical synthetic | |
|---|---|---|---|---|
| Method | made using recombinant DNA techniques, E. coli | (Recasens 2014, #1248) (?? PFF (?)/(7)) homogenization → sucrose gradient fractionation (→ 1)IF ii) filter retardation assay using antibodies against α-syn, p-aSyn, ubiquitin or p62 so this is called 'LB-containing extract' | made by chemical peptide synthesis | |
| pros | (Honguma 2016) if the original DNA sequence of α-syn is used for bacterial expression, ~70% of the protein is mistranslated, with cysteine at position 136 instead of tyrosine, resulting from a combination of codon usage and sequence context (Masuda et al., 2006). → 그 부정도 tt38 - TAT alterations should be used. Recombinant polypeptides typically are composed of only L-amino acids (i.e. non D-amino acids). ALN-HR0HR | To study PTM. May not be appropriate in applications where the secondary or tertiary structure is critical. | ||
| Product | The size limit for synthetic peptides is only about 20 to 50 amino acids, cf B-1 IL13 commercial product | |||
| parameter modelling A: aggregation | Recombinant α-syn readily assembles into amyloid like fibrils at high concentration, by shaking | |||
| neurotoxicity | ||||
| cytoskeleton damage | ||||
| receptor toxicity in vivo and cerebral amyloidosis | ||||
| synaptic plasticity disruption | ||||
| cognitive impairment in vivo and ex vivo | ||||
Skin aSyn
| content | |
|---|---|
| Syn / Syn-One test | https://cnedlifesciences.com/introducing-syn-one?utm_source=NMA%20Neurodegeneration_medium=Banner&utm_campaign=lazno%20Syn-One&utm_content=DORELLM30s%2k0 |
| Visual proof of phosphorylated α-syn co-located in dermal nerves of PD patients (~26) showed p-syn deposition (p < 0.001), Chi-square. | |
| (Donadio, 2024 #2752) | cutaneous phosphorylated α-synuclein detected by skin biopsy were 92.7% (89 of 96) with PD, 98.2% (54 of 55) with MSA, 96.0% (48 of 50) with DLB, and 100% (22 of 22) with PAF. |
Species
a-syn primary conformation
- The human alpha-syn protein is made of 140 amino acids, 14.46 kDa.
- Natively Unfolded, but becomes structured when bound to phospholipids
- (2019 Mende) α5 is likely to exist in vivo as an equilibrium mixture of unstructured monomer and statistically disfavoured helical oligomers, perhaps partially folded at membranes through phospholipid interactions
Primary aSyn species
| Pathologic conversion | Pathological aggregation | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| monomeric | tetramer | Phosphorylated d a-syn | Misfolded monomer | β-pleat sheet / β-sheet | insoluble | Oligomeric (still soluble) (Wang et al., 2016) | pre amyloid >220 nm | insoluble (amyloid-like) (long fibril) | Lewy bodies | ||
| (structure) | Overexpression of α-syn or of the mutant itself does not induce mass-like prefibrils in cells. The pathology was observed in the brains of mice injected with soluble human α-syn (degraded) | (Kim, 2018 #1244) Phosphorylation results show physiological α-syn exists as a folded helical tetramer that transition to monomers, under phosphorylation occurs at serine 129 of the helthy α-syn becomes insoluble | Phosphorylated α-syn at S87 5129 vs Y125 (Majbour 2018 #1255), and 90% of accumulated α-syn in LBs disease-related α-syn exists as a folded helical tetramer of α-syn at serine 129 of unhealthy (b) cytoplasm phosphor (b) S129 gradually become insoluble | β-sheet β-sheet | -Tong et al. 2010, Zhou et al. 2011, Murphy et al. 2014: misfolded forms of α-syn in PD brains whereas soluble forms are not significantly changed or even reduced (proteinase K-resistant α-syn-immunoreactivity of abnormal α-syn is enhanced by pretreatment with proteinase K (PK)) D α-syn aggregate D pSer129 a cysteine 3 (cysteine 그) signal-amplification of α-syn DG (PerkinElmer) (Croisier 2006 #2822) ↑ pSer129 α-syn aggregate (PerkinElmer) (Croisier 2006 #2822) ↑ pSer129 α-syn and its fraction in total α-syn ratios in the | (Rodriques, 2022 #2290) typically: a spheroid ~ 30 nm in diameters and 2 and 10 nm in height (Suchina, 2010 #2191) (X-ray ?) β-sheet (1-7 mm) (5/10/2017 일정) (RR Reviews) (PVR/Spheres 일정) Quantification 일 ?) hollow morphology | typically: a spheroid ~ 30 nm in diameters and 2 and 10 nm in height (Suchina, 2010 #2191) | (Rodriques, 2022 #2290) fibrils are stable organized molecules cool-like 7-13 nm in diameter extended PNAC domain in 1-43 (Pansson 2018 CDM2 cleared mass-was 85.4 MD2 ~5500 molecules on average (Mu-syn 14.46 kDa)) the lifespan of those length was 0.90 µm in [21]; ε in standard fix 0.94-1.5/0 (m diameter 0/0/2014, intra-stabilized by hydrogen-bonded β-strands perpendicular to the fiber axis and forming β-sheets but polymorphic and are often an association of protofibrils | |||
Uncertain Spans
| location | transcription | uncertainty |
|---|---|---|
| Sources / Brain extract / Method | (Recasens 2014, #1248) (?? PFF (?)/(7)) | reads as written; the placeholder notation is preserved verbatim. |