Parkin/PINK1 Mitophagy Evidence And pS65-Ub Detectability Notes

Parkin/PINK1 Mitophagy Evidence Matrix

Pathologic Row / Park2 Mutation

Row labels and effect-chain fragments:

pathologic
Park2 mutation
LOF
↓ pUb binding with Parkin (G284R, R275W)
abnormal protein folding & ↓ protein structural stability
(2019 Yi?) -> ↓ protein expression
↓ Ub? phosphorylation (R42P, V56E?) -> ↓ Parkin activation
↓ ubiquitination
↓ fission
↓ mitophagy
Mito rupture
↑ mt DNA release
↓ MC1 mass/activity
↓ ATP
↑ ROS
↑ Apoptosis
Neuronal loss

Upper-column fragment:

(mitochondria network) by Parkin's effect on DRP1

Extracellular Mitochondrial Expulsion Figure

Figure title:

Extracellular mito expulsion
(Choong, 2020 #1193)

Figure labels and numeric/statistical marks are too small to transcribe safely from this photo.

Evidence Rows

Evidence-row entries from the matrix:

Evidence type	Stable visible content
Postmortem human	아직 못 발견; Not found
Human CSF	cf Parkin
Human serum	Parkin can be detected in CSF & serum (Castellazzi, 2019 #891)
Mouse	(Palacino, 2004 #726): ↓ complex I & iv, ↓ ETC capacity, ↓ mito respiration, but normal morphology; (Noda, 2020 #701) KO Mouse, fragmented mito; (Palacino, 2004 #726): ↓ proteins re protection from OX; (Noda, 2020 #701) neuronal loss (2y)
iPS	(Suzuki, 2017 #680) ↑ ROS in park2 iPS; (Suzuki, 2017 #680) ↑ apoptosis (cleaved caspase3-positive cells (Fig. 3B).) in park2 iPS
fibroblast	아래 'fibroblast 참조'

Biomarker And Antibody Notes

Notes in the lower part of the matrix:

include LOF
Exclude Hyperactive Parkin variants (by known sequencing? Fibroblast?)
only MC1 PET+Ve patients?
 
Antibodies to PINK1 are being developed by MJFF (Padmanabhan, 2019 #820)
 
pS65 ub,
- Can we discriminate free pS65-ub and that bound to Parkin?
- Should be increased by PARKIN GT in non-neuronal cell type
  (그렇지만 gt roa 상 말초에서 변화 측정은 무의미하겠네)
 
pS65 Parkin antibody is being developed by MJFF (Padmanabhan, 2019 #820)
 
Antibody to activated Parkin is being by MJFF (Padmanabhan, 2019 #820)
 
Parkin substrate: VDAC1 (2018 Ordureau)
- Antibodies to Parkin substrates (VDAC, TOMM20, MFN2) are being developed by MJFF
  (Padmanabhan, 2019 #820)
 
Mitophagy in peripheral?
(fibroblast?, cf, mitophagy in heart tissue in mice, 2018 Sliter fig1)
 
BCPP MC1 imaging
 
(Geldenhuys, 2014 #733):

Parkin In sPD And pS65-Ub Detectability

Parkin in sPD row:

Parkin in sPD
 
postmortem
(Lonskaya, 2013 #1742) 12sPD,
(↑ phosphorylation -> ) ↓ (~50%, WB) Parkin level &
↓ soluble parkin, ↑ insoluble parkin (in striatum,) (not in cortex),
SN 은 안 본 듯

pS65-Ub reference text:

expression of pS65-ub
 
(Fiesel, 2015 #700): The increase of pS65-Ub upon mitochondrial damage was not
cell type specific since it occurred in several non-neuronal and neuronal-like
cells (Appendix Fig S3B).
 
some other references:
- Disease relevance of phosphorylated ubiquitin (p-S65-Ub)
  Fabienne C Fiesel and Wolfdieter Springer
- Sensitive ELISA-based detection method for the mitophagy marker p-S65-Ub
  in human cells, autopsy brain, and blood samples
  Jens O. Watzlawik

Detectability table entries:

Group	Barely detectible	Detectible
Fiesel, 2015	in young normal human (fig 7a, just one case 37세); HeLa cell (원래 no parkin); in human iNeurons from PINK1 patients; in PD patients' fibroblasts with a PINK1 mutation; PINK1 postmortem brain SN; In vitro: PINK1 멀쩡해도 PARKIN 없으면 적다 (2018 Ordureau fig 6, in vitro)	human: ↑ normal aged brain (SN, fig 7a, autopsy, just one case of 93 y); In vitro: ↑, mito stress in cells; can be induced in human iNeurons from controls; ↑ (greatly) functional parkin (method: HeLa cell overexpressing WT parkin ie 'HeLa Parkin') in HeLa cell (fig 2a, in the presence of CCCP, likely through enhanced formation of poly-Ub chains that in turn serve as substrates for PINK1...)

The bottom of the detectability table is cut by the photo, so the final phrase after PINK1 is incomplete.

Uncertain Spans

• 2019 Yi?, Ub? phosphorylation, R42P, and V56E? in the pathologic Park2 mutation row.
• Whether MC1 should be MCI, mitochondrial complex I, or another local shorthand in ↓ MC1 mass/activity and BCPP MC1 imaging.
• Human Serum is visible as Huan Serum in OCR; interpreted as Human Serum.
• The exact column alignment of 아직 못 발견, Not found, dashes, and evidence rows across the matrix is uncertain because the table is extremely wide and photographed at an angle.
• Antibody to activated Parkin is being by MJFF appears grammatically incomplete but is transcribed as visible.
• only MC1 PET+Ve patients? may mean PET+ve or another local notation.
• gt roa in the Korean note is preserved as visible shorthand; exact intended expansion is unclear.
• The pS65-Ub detectability table is cut at the bottom; the final red note after for PINK1 is incomplete.