BACE1- and BACE2-expressing Human Cells

Andrau, D.; Dumanchin-Njock, Cécile; Ayral, Erwan; Vizzavona, Jean; Farzan, Michael; Boisbrun, Michel; Fulcrand, Pierre; Hernandez, Jean‐François; Martinez, Jean; Lefranc-Jullien, S.; Checler, Frédéric

doi:10.1074/jbc.m302622200

Cited by 72 publications

(37 citation statements)

References 72 publications

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“…To investigate BACE activity in TMEM59 expressing cells a fluorimetric assay (48,49) was used. HEK293 cells were cotransfected with BACE1 and either control plasmid or TMEM59.…”

Section: Methodsmentioning

confidence: 99%

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The Novel Membrane Protein TMEM59 Modulates Complex Glycosylation, Cell Surface Expression, and Secretion of the Amyloid Precursor Protein

Ullrich

Münch

Neumann

et al. 2010

Journal of Biological Chemistry

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Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases ␣-and ␤-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid ␤ peptide (A␤). At present, little is known about the cellular mechanisms that control APP shedding and A␤ generation. Here, we identified a novel protein, transmembrane protein 59 (TMEM59), as a new modulator of APP shedding. TMEM59 was found to be a ubiquitously expressed, Golgi-localized protein. TMEM59 transfection inhibited complex N-and O-glycosylation of APP in cultured cells. Additionally, TMEM59 induced APP retention in the Golgi and inhibited A␤ generation as well as APP cleavage by ␣-and ␤-secretase cleavage, which occur at the plasma membrane and in the endosomes, respectively. Moreover, TMEM59 inhibited the complex N-glycosylation of the prion protein, suggesting a more general modulation of Golgi glycosylation reactions. Importantly, TMEM59 did not affect the secretion of soluble proteins or the ␣-secretase like shedding of tumor necrosis factor ␣, demonstrating that TMEM59 did not disturb the general Golgi function. The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. Both proteins are required for normal localization and activity of Golgi glycosylation enzymes. In summary, this study shows that TMEM59 expression modulates complex N-and O-glycosylation and suggests that TMEM59 affects APP shedding by reducing access of APP to the cellular compartments, where it is normally cleaved by ␣-and ␤-secretase. Processing of the amyloid precursor protein (APP)3 by two different proteases, called ␣-and ␤-secretase, is a central regulatory event in the generation of the amyloid ␤ peptide (A␤), which has a key role in Alzheimer disease (AD) pathogenesis (1). Both ␣-and ␤-secretase cleave the type I membrane protein APP within its ectodomain close to its transmembrane domain (2). This leads to the secretion of soluble forms of APP (APPs) and is referred to as APP shedding. The ␤-secretase is the aspartyl protease BACE1 and cleaves APP at the N terminus of the A␤ peptide domain, thus catalyzing the first step in A␤ peptide generation (3, 4). After the initial cleavage of APP by BACE1, the remaining C-terminal APP fragment is cleaved by ␥-secretase within its transmembrane domain at the C terminus of the A␤ domain, leading to the secretion of the A␤-peptide (5). In contrast to ␤-secretase, ␣-secretase cleaves within the A␤-sequence of APP, and thereby precludes A␤ peptide generation. Additionally, ␣-but not ␤-secretase cleavage generates a secreted form of APP (APPs␣), which has neurotrophic and neuroprotective properties (6 -8). The ␣-secretase is a member of the ADAM family (A disintegrin and metalloprotease) of proteases (9 -12). At present little is known about how the cell controls access of APP to its secretases and the amount of ␣-and ␤-secretase cleavage (reviewed in Refs. 13 and 14). Recent studies increasingl...

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“…To investigate BACE activity in TMEM59 expressing cells a fluorimetric assay (48,49) was used. HEK293 cells were cotransfected with BACE1 and either control plasmid or TMEM59.…”

Section: Methodsmentioning

confidence: 99%

“…TMEM59 expression did not affect the activity of ␣-and ␤-secretase, as measured by established assays using fluorogenic substrates (46,48) (Fig. 2, G and H).…”

Section: Tmem59 Modulates Maturation and Shedding Of App-tomentioning

confidence: 99%

The Novel Membrane Protein TMEM59 Modulates Complex Glycosylation, Cell Surface Expression, and Secretion of the Amyloid Precursor Protein

Ullrich

Münch

Neumann

et al. 2010

Journal of Biological Chemistry

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“…BACE1 and ADAM10 Fluorimetric Assay-Cells were lysed with 10 mM Tris-HCl, pH 7.5, and then homogenates were monitored for their BACE1 activity as described previously (39,42). Briefly, samples (30 g of proteins diluted in 10 mM acetate buffer, pH 4.5) were incubated in a final volume of 100 l of the above acetate buffer containing BACE1 substrate (10 M, (7-methoxycoumarin-4-yl)acetyl-SEVNLDAEFRK(2,4-dinitrophenyl)-RRNH2; R & D Systems) in the absence or presence of the previously described BACE1 inhibitor JMV2764 (50 M) (39).…”

Section: Cell Cultures and Transfections-fibroblasts Devoid Of Ps1mentioning

confidence: 99%

NFκB-dependent Control of BACE1 Promoter Transactivation by Aβ42

Buggia-Prévot

Sévalle

Rößner

et al. 2008

Journal of Biological Chemistry

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111

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␤-Amyloid (A␤) peptides that accumulate in Alzheimer disease are generated from the ␤-amyloid precursor protein (␤APP) by cleavages by ␤-secretase BACE1 and by presenilindependent ␥-secretase activities. Very few data document a putative cross-talk between these proteases and the regulatory mechanisms underlying such interaction. We show that presenilin deficiency lowers BACE1 maturation and affects both BACE1 activity and promoter transactivation. The specific ␥-secretase inhibitor DFK167 triggers the decrease of BACE1 activity in wild-type but not in presenilin-deficient fibroblasts. This decrease is also elicited by catalytically inactive ␥-secretase. The overexpression of APP intracellular domain (AICD), the ␥/⑀-secretase-derived C-terminal product of ␤-amyloid precursor protein, does not modulate BACE1 activity or promoter transactivation in fibroblasts and does not alter BACE1 expression in AICD transgenic brains of mice. A DFK167-sensitive increase of BACE1 activity is observed in cells overexpressing APP⑀ (the N-terminal product of ␤APP generated by ⑀-secretase cleavage harboring the A␤ domain but lacking the AICD sequence), suggesting that the production of A␤ could account for the modulation of BACE1. Accordingly, we show that HEK293 cells overexpressing wild-type ␤APP exhibit a DFK167-sensitive increase in BACE1 promoter transactivation that is increased by the A␤-potentiating Swedish mutation. This effect was mimicked by exogenous application of A␤42 but not A␤40 or by transient transfection of cDNA encoding A␤42 sequence. The I B kinase inhibitor BMS345541 prevents A␤-induced BACE1 promoter transactivation suggesting that NF B could mediate this A␤-associated phenotype. Accordingly, the overexpression of wild-type or Swedish mutated ␤APP does not modify the transactivation of BACE1 promoter constructs lacking NF B-responsive element. Furthermore, APP/␤-amyloid precursor protein-like protein deficiency does not affect BACE1 activity and expression. Overall, these data suggest that physiological levels of endogenous A␤ are not sufficient per se to modulate BACE1 promoter transactivation but that exacerbated A␤ production linked to wild-type or Swedish mutated ␤APP overexpression modulates BACE1 promoter transactivation and activity via an NF B-dependent pathway.Alzheimer disease (AD) 3 is characterized by abnormal deposition of a set of hydrophobic peptides called amyloid ␤ (A␤) peptides. The increase of cerebral A␤ levels is one of the common denominators characterizing both sporadic and familial forms of AD and therefore, if not demonstrated as the etiological cause of AD pathology, is often considered as a key factor contributing to the degenerative process (1).The mechanisms by which A␤ peptides are generated are a matter of intense research in the AD field. A␤ peptides are released from a transmembrane protein, ␤-amyloid precursor protein (␤APP), by the sequential attacks by ␤-and ␥-secretase that liberate the N-and C-terminal moieties of A␤ peptides, respectively (2). All ␤-secretase-like activi...

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“…Thus, BACE likely fulfills some natural functional roles in HEK 293 cells. Additionally, HEK 293 cells were used previously to identify BACE and to analyze its proteolytic function by many other reported studies [27,28,30,31]. After transfection, all cells were grown in the culture media containing 0.5 mg/ml G418.…”

Section: Overexpression Of Bace and Bace2 In Cells And Bace Activity mentioning

confidence: 99%

“…BACE and a closely related enzyme BACE2 are two newly identified proteases which are involved in proteolysis of amyloid precursor protein (APP) [28][29][30][31]. BACE is a recognized enzyme that initiates generation of beta amyloid peptides, the major components of amyloid plaques found in postmortem brains of Alzheimer's disease patients.…”

Section: Introductionmentioning

confidence: 99%