Cathepsin D displays in vitro β-secretase-like specificity

Chevallier, Nathalie; Vizzavona, Jean; Marambaud, Philippe; Baur, Claus Peter; Spillantini, Maria Grazia; Fulcrand, Pierre; Martinez, Jean; Goedert, Michel; Vincent, Jean‐Pierre; Checler, Frédéric

doi:10.1016/s0006-8993(96)01330-3

Cited by 72 publications

(50 citation statements)

References 33 publications

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“…Although our data do not address the identity of specific lysosomal hydrolases(s) that may be involved in APP processing, one candidate is the aspartyl protease Cat D, which is trafficked by the mannose 6-phosphate motif (46), has ␤-secretase activity in some systems (8,(53)(54)(55), and has a genetic polymorphism that has been linked in some studies to increased risk for AD (56,57), although not all studies have found this linkage (58 -61). Our previous studies have shown that the neuronal expression, trafficking, and intracellular distribution of Cat D are significantly abnormal in AD (9,11,12), although these studies typically employed Cat D as a marker for more generalized changes in lysosomal hydrolase expression and intracellular distribution, as did the current study.…”

Section: Discussionmentioning

confidence: 89%

Alzheimer's Disease-related Overexpression of the Cation-dependent Mannose 6-Phosphate Receptor Increases Aβ Secretion

Mathews

Guerra

Jiang

et al. 2002

Journal of Biological Chemistry

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Prominent endosomal and lysosomal changes are an invariant feature of neurons in sporadic Alzheimer's disease (AD). These changes include increased levels of lysosomal hydrolases in early endosomes and increased expression of the cation-dependent mannose 6-phosphate receptor (CD-MPR), which is partially localized to early endosomes. To determine whether AD-associated redistribution of lysosomal hydrolases resulting from changes in CD-MPR expression affects amyloid precursor protein (APP) processing, we stably transfected APP-overexpressing murine L cells with human CD-MPR. As controls for these cells, we also expressed CD-MPR trafficking mutants that either localize to the plasma membrane (CD-MPRpm) or to early endosomes (CD-MPRendo). Expression of CD-MPR resulted in a partial redistribution of a representative lysosomal hydrolase, cathepsin D, to early endosomal compartments. Turnover of APP and secretion of sAPP␣ and sAPP␤ were not altered by overexpression of any of the CD-MPR constructs. However, secretion of both human A␤40 and A␤42 into the growth media nearly tripled in CD-MPR-and CD-MPRendo-expressing cells when compared with parental or CD-MPRpm-expressing cells. Comparable increases were confirmed for endogenous mouse A␤40 in L cells expressing these CD-MPR constructs but not overexpressing human APP. These data suggest that redistribution of lysosomal hydrolases to early endocytic compartments mediated by increased expression of the CD-MPR may represent a potentially pathogenic mechanism for accelerating A␤ generation in sporadic AD, where the mechanism of amyloidogenesis is unknown.Deposition in the brain parenchyma and cerebral vessel walls of ␤-amyloid and intracellular neurofibrillary tangles are diagnostic hallmarks of AD.1 ␤-Amyloid consists primarily of the short peptide A␤, which is derived from the proteolytic cleavage of the amyloid precursor protein (APP) at a site within its luminal domain (the ␤-cleavage site) and cleavage within the transmembrane domain (the ␥-cleavage site). Additionally, ␣-cleavage of APP may occur within the A␤ domain of APP, adjacent to the plasma membrane (see Ref.1 for a review of APP proteolytic processing). ␤-Amyloidogenesis in familial AD caused by mutation of APP or the presenilins involves, at least in part, the overproduction of A␤ (2); however, the mechanisms promoting ␤-amyloidogenesis in sporadic forms of AD, which account for Ͼ90% of AD cases, remain unclear. Experimental evidence has shown that early endosomes are an important site for APP processing, including the generation of A␤. Expression of trafficking mutants of APP lacking endocytosis signals reduces A␤ production compared with endocytosis-competent wild-type APP (3, 4). There is also evidence that at least some of the proteases responsible for A␤ generation reside within early endosomes. For example BACE, a recently identified transmembrane aspartic protease with ␤-secretase activity, resides in part within early endosomes (5-7). The lysosomal protease cathepsin D (Cat D) was recently confirmed ...

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Section: Discussionmentioning

confidence: 89%

Alzheimer's Disease-related Overexpression of the Cation-dependent Mannose 6-Phosphate Receptor Increases Aβ Secretion

Mathews

Guerra

Jiang

et al. 2002

Journal of Biological Chemistry

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“…7B). We took advantage of the description of cathepsin D as an in vitro ␤-secretase-like activity (40,41) to further compare the usefulness and accuracy of these two assays. Interestingly, although purified cathepsin D potently hydrolyzed CS and JMV2235, this enzyme was unable to hydrolyze the mutated substrate JMV2236 (Fig.…”

Section: Comparison Of Jmv-based Assay With a Commercial ␤-Secretase mentioning

confidence: 99%

“…Thus, cathepsin D, a protease with in vitro-like ␤-secretase activity (40,41), indeed cleaves efficiently a commercial substrate mimicking the ␤-secretase-targeted sequence. However, our assay demonstrated that this protease did not behave as a good ␤-secretase candidate because it did not hydrolyze JMV2236, the fluorimetric substrate bearing the Swedish mutation.…”

Section: ␤-Secretase In Bace1-and Bace2-expressing Human Cellsmentioning

confidence: 99%

BACE1- and BACE2-expressing Human Cells

Andrau

Dumanchin-Njock

Ayral

et al. 2003

Journal of Biological Chemistry

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We have set up stably transfected HEK293 cells overexpressing the ␤-secretases BACE1 and BACE2 either alone or in combination with wild-type ␤-amyloid precursor protein (␤APP). The characterization of the ␤APP-derived catabolites indicates that cells expressing BACEs produce less genuine A␤1-40/42 but higher amounts of secreted sAPP␤ and N-terminal-truncated A␤ species. This was accompanied by a concomitant modulation of the C-terminal counterpart products C89 and C79 for BACE1 and BACE2, respectively. These cells were used to set up a novel BACE assay based on two quenched fluorimetric substrates mimicking the wild-type (JMV2235) and Swedish-mutated (JMV2236) ␤APP sequences targeted by BACE activities. We show that BACEs activities are enhanced by the Swedish mutation and maximal at pH 4.5. The specificity of this double assay for genuine ␤-secretase activity was demonstrated by means of cathepsin D, a "false positive" BACE candidate. Thus, cathepsin D was unable to cleave preferentially the JMV2236-mutated substrate. The selectivity of the assay was also emphasized by the lack of JMV cleavage triggered by other "secretases" candidates such as ADAM10 (A disintegrin and metalloprotease 10), tumor necrosis ␣-converting enzyme, and presenilins 1 and 2. Finally, the assay was used to screen for putative in vitro BACE inhibitors. We identified a series of statine-derived sequences that dose-dependently inhibited BACE1 and BACE2 activities with IC 50 in the micromolar range, some of which displaying selectivity for either BACE1 or BACE2.

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“…Cathepsin D, an aspartyl protease, was initially implicated as a ␤-secretase candidate (Brown et al, 1996;Chevallier et al, 1997), although reduction in cathepsin D activity never effectively lowered A␤ levels (Saftig et al, 1996). Likewise, overexpression of cathepsin S resulted in enhanced A␤ secretion (Munger et al, 1995).…”

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confidence: 99%

Cathepsins B and L Differentially Regulate Amyloid Precursor Protein Processing

Klein¹,

Felsenstein²,

Brenneman³

2008

J Pharmacol Exp Ther

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Previous studies have shown that cathepsins control amyloid beta (A␤) levels in chromaffin cells via a regulated secretory pathway. In the present study, this concept was extended to investigations in primary hippocampal neurons to test whether A␤ release was coregulated by cathepsins and electrical activity, proposed components of a regulated secretory pathway. Inhibition of cathepsin B (catB) activity with CA074Me or attenuation of catB expression through small interfering RNA produced decreases in A␤ release, similar to levels produced with suppression of ␤-site APP-cleaving enzyme 1 (BACE1) expression. To test whether the catB-dependent release of A␤ was linked to ongoing electrical activity, neurons were treated with tetrodotoxin (TTX) and CA074Me. These comparisons demonstrated no additivity between decreases in A␤ release produced by TTX and CA074Me. In contrast, pharmacological inhibition of cathepsin L (catL) selectively elevated A␤42 levels but not A␤40 or total A␤. Mechanistic studies measuring C-terminal fragments of amyloid precursor protein (APP) suggested that catL elevated ␣-secretase activity, thereby suppressing A␤42 levels. The mechanism of catB-mediated regulation of A␤ release remains unclear but may involve elevation of ␤-secretase. In summary, these studies provide evidence for a significant alternative pathway for APP processing that involves catB and activity-dependent release of A␤ in a regulated secretory pathway for primary neurons.

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Cathepsin D displays in vitro β-secretase-like specificity

Cited by 72 publications

References 33 publications

Alzheimer's Disease-related Overexpression of the Cation-dependent Mannose 6-Phosphate Receptor Increases Aβ Secretion

Alzheimer's Disease-related Overexpression of the Cation-dependent Mannose 6-Phosphate Receptor Increases Aβ Secretion

BACE1- and BACE2-expressing Human Cells

Cathepsins B and L Differentially Regulate Amyloid Precursor Protein Processing

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