| multi-vesicular endosomes | multivesicular bodies (MVB) | ||
|---|---|---|---|
| dynamic tubular-vesicular network (vesicles up to 1 μm in diameter with connected tubules of approx. 50 nm diameter). | Multivesicular appearance, mainly spherical, lack tubules, and contain many close-packed intraluminal vesicles. | ||
| Marker | Markers include RAB5A and RAB4, Transferrin and its receptor and EEA1 | Markers include RAB7, RAB9, and mannose 6-phosphate receptors.[10] | |
| 3. use with the plasma membrane instead of with lysosomes, | |||
| releasing the lumenal vesicles, now called exosomes, into the extracellular medium. |
Macroautophagy
Autophagy markers
xxix) resource
a. https://www.novusbio.com/support/faqs-autophagy-and-lc3 (very good)
The human LC3 gene family has three members, LC3A, LC3B and LC3C, The form LC3-II is one of the main components of the autophagosome membrane (LC3A-II and LC3B-II, also LC3C-II but not studied here) that resides in both the inner and outer site of the membrane. The LC3-II is derived from a proLC3 ~30KDa protein after cleavage by autophagin Atg4 to produce the active cytosolic form LC3-I. This in turn is activated by Atg7, and then transferred to Atg3, a second E2-like enzyme, becoming a membrane-bound form, LC3-II [5]. After autophagosome formation, the LC3-II located in the outer site is released to the cytosol and the LC3-II located in the inner site is degraded by hydrolases [6]. In this latter form, LC3-II localizes on the spherical autophagosomal and autolysosomal membranes, forming a suitable marker of autophagic activity [6,7].
아래 각각 존재함
LC3A-I, LC3A-II,
a. LC3B-I, LC3B-II (I used these two)
Microautophagy
xxx) the sequestration of proteins directly into lysosomes by invagination of the lysosome membrane and subsequent degradation in the lysosomal lumen (
CMA
Process
- a highly specific subset of cytosolic proteins are recognised by heat-shock cognate 70 (HSC70) on their pentapeptide recognition motif, KFERQ, → transportation directly into the lysosomes via interaction with LAMP2A (Lynch-Day et al., 2012, Cuervo et al., 2004b).
- cf) HSC70 is a cytosolic member of the Hsp70 chaperone family,5
Substrates
- only cytosolic proteins bearing a KFERQ-like motif represent CMA substrates
- aSyn in neurons
- disease-associated mutant A30P or A53T a-Syn displays higher binding affinity to Lamp2a, and “cloggs” the CMA translocation channels, contributing to neurotoxicity
Pathology
- (CMA activity depends on Lamp2a and Hsc70 levels) these two proteins are decreased in PD human postmortem brain material.10-14
Cf)
| HSP70 Family | ||
| HSC70 | HSP70 | |
| constitutively expressed | Stress-induced | |
| synonym | Hsp73/HSPA8 | |
BMP (Bis(monoacylglycero)phosphate)
-
What is it?
- a negatively charged (anionic) glycerophospholipid
-
Presence
- could reach up to 30-60 mol% of the total phospholipids in the lysosomal internal membranes (13, 20).
- Many cell types have low amounts of BMP, which comprises less than 1% of total cellular phospholipid content
- BMP is primarily enriched in endosomal/Lysosomal membranes (di22:6 according to Denali R&D Day)
-
Function
- BMP conveys negative surface charge to the luminal lysosomal vesicles, even at pH lower than 5 (21).
- Stimulate GlcCer degradation by GBA1 (by facilitating interaction of hydrolases and activator proteins with sphingolipid substrates) (by mediating electrostatic interactions with positively charged saposins and lysosomal lipases (e.g., GBA).)
- (1) digestion and recycling of membranous materials, (2) regulation of cholesterol homeostasis, (3) trafficking of lysosomal enzymes, and (4) formation of intra luminal vesicles (Schmitz and Müller, 1991).
-
BMP change in diseases
| BMP | |||
|---|---|---|---|
| BMP | (GBA) PD | Human Urine | (Alcalay, 2020 #649) 원래 LRRK2환자 분석인데 GBA도 있음, 표 왼쪽 보면 (검은점들) 다소 GBA+PD+LRRK2- vs GBA-PD+LRRK2- 의 차이 별로 없는 듯 한데? |
| LSDs | (Akgoc, 2015 #648) ↑ BMP in many LSDs, including mu copolysaccharidosis, Niemann-Pick disease type A/B/C, Gaucher disease (막상 아래 Hein 의 in vitro study 말고는 안 보임) and Fabry disease ( 15-26 ). Our results showed that the content of BMP in brain was significantly greater in humans and in animals (mice, cats, American bears) with either GM1 or GM2 ganglioside storage diseases, than in brains of normal subjects. BMP was elevated in tissue of a SD (Sandhoff disease) human patient, and in serum obtained from GM2 gangliosidoses patients ( 20, 27 ). However, detailed study of BMP content in the GM2 and GM1 gangliosidoses brain is lacking ( 28 ). | ||
| (Hein, 2013 #670) Mechanism of ↑ BMP in LSD: expansion of the lysosomal network → increase in BMP | |||
| GD | In vitro | (Hein, 2013 #670) ↑ BMP in THP-1 macrophage model of GD | |
| GRN-FTD | Human CSF | (Denali: PGRN_KSQ_Apr2020Update) two BMP species are decreased in the CSF of GRN-FTD patients as well as in that of a subset of non-GRN FTD patients: BMP36:2 (18:1/18:1) and BMP44:12 (22:6/22:6). - PTV:PGRN PRC1 slide: Denali plans to examine CSF BMP of FTD patients | |
| What model? | In vivo brain | BMP is increased in mice model Denali R&D Day) LRRK2 inhibitors (→ ↓ BMP Di22:6 | |
| (Inline Takeda slide titled "ETV:IDS (DNL310) RESCUES BRAIN BMP LEVELS · Brain BMP" — di-22:6-BMP levels (ng/mg protein) bar chart over groups Vehicle (TfRmu/hu), Vehicle / IDS / ETV:IDS (IDS KO; TfRmu/hu) Multi-dose. *** and ** significance brackets between vehicle vs ETV:IDS and IDS vs ETV:IDS. Preserved as body_r03 evidence.) | |||
| 현재로서 disease에서 BMP가 ↑ 인지 ↓ 인지조차 불확실함. Lysosomal dysfunction 의 원인인지, 결과인지도 불명. | |||
| GBA PD | Human CSF & plasma | Takeda DMPK: BMP measurement ongoing | |
| GBA PD | Human postmortem | Not available? 해야 되지 않나? | |
BMP Correction
| healthy people | Urine | Denali R&D Day) LRRK2 inhibitors → ↓ BMP |
| In healthy human : CSF | Denali R&D Day) LRRK2 inhibitors (for 10 dyas dosing) → ↓ BMP Di22:6 | |
| animal model | In vivo brain & CSF | Denali R&D Day) LRRK2 inhibitors (→ ↓ BMP Di22:6 mice |
(Inline Takeda slide titled “ETV:IDS (DNL310) RESCUES BRAIN BMP LEVELS · Brain BMP” repeated; preserved as body_r05 evidence.)
Uncertain Spans
| location | transcription | uncertainty |
|---|---|---|
| BMP urine row | Alcalay, 2020 #649 Korean fragment 다소 GBA+PD+LRRK2- vs GBA-PD+LRRK2- | The boolean qualifier LRRK2- could read as LRRK2+ depending on small superscript symbol. |
BMP Akgoc, 2015 #648 row | mu copolysaccharidosis | Likely “mucopolysaccharidoses”; transcribed as the source spells it. |
| ETV:IDS slide group labels | IDS KO; TfRmu/hu | The mu/hu superscript glyphs are tiny; “mu/hu” is read as in the slide caption. |
| BMP Correction row | for 10 dyas dosing | Source typo; “dyas” is intentionally preserved as written. |