총정리 GBA protein & activity (continued), Sidransky 2012 review, Saposin C summary

source	visible cells
Gegg 2015	in areas of the brain with low asyn pathology 라고 언급있을 뿐 그 level은 언급 없음.
Gegg 2012	↓ gba activity & expression(?) in PD; adjacent cell: a-syn or mitochondrial dysfunction → ↓ GBA activity
Parkkinen 2011	PD + heterozygous GBA mutation VS PD only; adjacent cell: No difference in the number of LB aggregates; note: Normal 과 비교한 게 아니니까 당연한 결과 아닌가?
Murphy

[Gegg 2012] GBA activity / protein level

context	GBA Activity	GBA protein level (BCA가 아니라 모두 WB로 했네?)	note / conclusion
In human PD + hetero GBA mut	↓ in all brain areas (58% SN, except frontal cx)	↓	PD pathology is exacerbated and accelerated, but not necessarily initiated, by GBA mutations (Fig 5). This would explain why not all GBA mutation carriers develop PD, and why those who do tend to do so at an earlier age.
In human PD + no GBA mut	↓ SN (33%) and cbll	↓	**GBA mutation없는데도 ↓ GBA level & activity 이네!!
SH-SY5Y that lacks PINK1 (resulting in mito dysfunction & OS)	↓	↓	이 mechanism은 모르겠다 함.

Table Below by Sidransky 2012 review

Author	Model	Result	Criticism
Cleeter (2013)	(undifferentiated) 1. SH-SY5Y ++CBE 2. SH-SY5Y w/o GBA	1. ↓ Mito functio 2. ↑ a-syn 3. no change in lysosomal 4. no change in UPS
V Cullen, SP Sardi, J Ng et al. Acid β-glucosidase mutants linked to Gaucher disease, Parkinson disease, and Lewy body dementia alter alpha-syn processing Ann Neurol, 69 (2011), pp. 940-953	Cell lines MES	↑ a-syn	thus not supporting loss-of-function theory)
	PC12 w CBE (10,50,100uM), duration not said	No change
	HEK293	a-syn altered
	Mouse (a point muta KI) /D409/V/D09V	↑ a-syn in hippocampus (by both mutation & enz downregulation) that could be reversed by rapamycin, an inducer of macroautophagy Not in heterozygote mouse	D409V mut is not reported in GD and doesn't manifest typical Gaucher features. The neuropathology in the mice suggest that a-syn is not associated with GBA activity or substrate accumulation (이 두 비판은 Sidransky's review 2012)
SP Sardi, J Clarke, C Kinnecom et al. CNS expression of glucocerebrosidase corrects alpha-syn pathology and memory in a mouse model of Gaucher-related synopathy Proc Natl Acad Sci USA, 108 (2011), pp. 12101-12106	Mouse (a point muta KI) /D409/V/D09V	↑ a-syn in hippocampus ↓ memory (reversed by GBA)
AB Manning-Bog (US), Schule, JW Langston alpha-syn-glucocerebrosidase interactions in pharmacological Gaucher models: a biological link between Gaucher disease and parkinsonism Neurotoxicology, 30 (2009), pp. 1127-1132	1. Cell line/ 2. Ventral mesencephalon from WT Mouse treated with CBE	↑ monomeric a-syn 근데 RT-PCR에선 no change 즉 expression증가가 아니라 clearance감소가 원인이겠구나..
H Xu, L Jia, B Quinn et al. global gene expression profile progression in Gaucher disease mouse models BMC Genomics, 12 (2011), p. 20	Mouse / homozygous for V394L or D409H, which have a normal lifespan and do not show storage of glucosylceramide in the CNS, were crossed with mice deficient in prosaposin, including the peptide saposin C,	↑ a-syn in cortical neurons (이상하다 이 논문에 a-syn 없는뎅? 2008논문에도 없고)	Model 은 괜찮으나 (정상수명?이라는 점과 기질축적이 없음이라는 단점을 saposin C로 보상했으니) a-syn aggregation was not related to glucosylceramide accumulation 원래 이 모델은 기질 축적이 없는데 saposin 없애서 기질 축적을 유도했음. (원래 saposin은 sphingolipid분해에 역할)
		a-syn and GBA bind physically in normal lysosomes and statilize a-syn to prevent degradation

Saposin C summary

source / year	model	note
1971	In vitro	GD 환자로부터의 spleen 과 정상것 섞었더니 GBA activity회복. (GBA amount난 size는 아님), 의미: GBA deficiency에서 compensation으로 이것이 이루어진다는 것으로 해석될 수 있는 것은 이게 유일.
Prence / 1985	in vitro	saposin C and/or PS : ↑ GBA (in activity & size from 57->188kDa). it appears that saposin promotes the transfer of PS. 그러나 이 근거는 본문에 못 찾겠음. 이말이 (in vivo) cell memb에 있는 PS를 within lysosome으로 옮긴다는 말은 아니겠군.
1985	in vitro	Saposin C activates GBA for both its natural substrate and artifical substrate (ie 4muglc). Saposin C binds to GBA (but not the substrate).
		PS 도 기전이 gba 에 binding 하는 것이라고 읽힘. (but different sites)
Datta / 1986	in vitro (N1E-115) 'Uptake by neuroblastoma cells'	Glucysylceramide → ↑ the amount of Saposin C, measured by immunoassay
1986 / 'Glucosylceramide and the Level of the Glucosidase-Stimulating Proteins'	mouse	CBE 100mg/kg ip single (겨우 single이니까 내거랑 다를 수 있겠음) → ↑ the amount of Saposin C by 60-70%, persisting at least 7 days, measured by immunoassay / draw the picture below. GBA 의 size아니고 activity임.
1988	MOUSE	Phosphatidylserine → ↓ liver weight, ↓ liver lipid, ↓ glucosylceramide

Lower Saposin / GBA table

row	prosaposin	Saposin C	GBA (unactivated)	GBA (activated)
amino acid	524	80
molecular weight	70kDa	9kDa	60 kDa	Aerts : 200 kDa (by HPLC), In the denatured state both forms of glucocerebrosidase reacted with anti-(placental glucocerebrosidase) antibodies We propose that form I glucocerebrosidase is a monomeric form of the enzyme, whereas form I1 glucocerebrosidase is a high-M, complex of the enzyme in association with sphingolipidactivator protein 2.(=saposin C) 이게 SC와 결합되어 있는 거라고 말함? Prence: 188 kDa when added with Saposin C & PS (by Sucrose gradient), 이건 GBA자체가 커지는 거라고 읽히고.
lower note	WB엔 이게 잡히는 거고	GBA와 complex는 이게 만들고

Saposin C deficiency → GD like feature 이건 증거 있는 것 같음. … Saposin C가 accumulation 되어 있다는 얘기가 Datta: uptake 의 introduction에 라는 데 원본을 찾아봐야 하나?

Uncertain Spans

location	transcription	uncertainty
top continuation	the Gegg 2015 / Gegg 2012 / Parkkinen 2011 / Murphy rows continue from prior captures’ wider matrix; full column headers are above the visible crop.	The 2-column collapse here is intentional because the upstream column structure is not re-shown in this photo.
Sidransky table / Cleeter row	↓ Mito functio	reads as written; the trailing n is clipped from function.
Sidransky table / Cullen row	D409/V/D09V; (thus not supporting loss-of-function theory)	the genotype string is visually/OCR uncertain and likely D409V/D409V; the closing parenthesis on the criticism cell has no matching open paren in the visible text.
Sidransky table / Manning-Bog row	author/title/citation	the leftmost column is partially clipped at the photo edge; visible portion preserved.
Saposin summary	source names for several rows (1971, 1985, second 1986, 1988)	The leftmost source name column is clipped at the photo edge; only year markers are visible.
Saposin summary	artifical substrate, 4muglc, Glucysylceramide	reads as written; likely authoring/OCR slips for artificial, 4muGlc (4-methylumbelliferyl-β-D-glucopyranoside), and Glucosylceramide.