Characterization by radiosequencing of the carboxyl-terminal derivatives produced from normal and mutant amyloid β protein precursors

Cheung, Tobun T.; Ghiso, Jorge; Shojif, Mikio; Cai, Xiao Dan; Golde, Todd E.; Gandy, Samuel E.; Frangione, B; Younkin, Steven G.

doi:10.3109/13506129409148622

Cited by 25 publications

(16 citation statements)

References 25 publications

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“…These results suggest that the secretory processing of APP may be heterogenous in DS brain, although some effects of post-mortem proteolysis cannot be completely ruled out. However, similar results to ours have been reported recently by Zhong et al and Cheung et al [26,27] in culture cells, who noted cleavage of APP near the membrane surface at several different sites. Additionally, a C-terminal fragment starting at residue -30 (A/3 numbering) has been reported in Fig.…”

Section: Resultssupporting

confidence: 66%

Secretory cleavage site of Alzheimer amyloid precursor protein is heterogeneous in Down's syndrome brain

et al. 1994

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A~ ~/A4) is the major constituent of brain amyloid in Alzheimer's disease (AD), Down's syndrome (DS) and normal aged persons. This protein is presumably derived by normal proteolysis from a precursor protein (APP). In this study, C-terminal fragments of APP in a Tris/Triton soluble fraction were partially purified from DS brain by heparin-affinity and reverse phase chromatography, and analyzed by N-terminal amino acid sequencing after SDS polyacrylamide gel electrophoresis and Western blotting. We found at least six different C-terminal fragments including those with the entire ,Aft region. These results suggest that secretory processing of APP is heterogeneous and generates amyloidogenic C-terminal fragments.

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Section: Resultssupporting

confidence: 66%

Secretory cleavage site of Alzheimer amyloid precursor protein is heterogeneous in Down's syndrome brain

et al. 1994

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“…Of note, it was frequently possible to detect a slowly migrating APP C-terminal fragment (Fig. 1B) potentially consistent with a "␤-C-terminal-fragment" or "C100-fragment" bearing A␤ at its N terminus (26) and thus constituting the immediate precursor of A␤. Immunochemical and radiochemical characterization of this APP C-terminal fragment is underway.…”

Section: ␣-Secretase Cleavage Of Human App Expressed In Yeast: Blockamentioning

confidence: 82%

Involvement of Cell Surface Glycosyl-phosphatidylinositol-linked Aspartyl Proteases in α-Secretase-type Cleavage and Ectodomain Solubilization of Human Alzheimer β-Amyloid Precursor Protein in Yeast

Komano

Seeger

Gandy

et al. 1998

Journal of Biological Chemistry

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Human ␤-amyloid precursor protein (APP) introduced into yeast undergoes ␣-secretase-type cleavage, suggesting that yeast have ␣-secretase-like protease(s). Here we report that two structurally and functionally related glycosyl-phosphatidylinositol-linked yeast aspartyl proteases, Mkc7p and Yap3p (collectively termed yapsin), are responsible for ␣-secretase-type cleavage of APP expressed in yeast, resulting in release of soluble APP into the extracellular space. Disruption of MKC7 and YAP3 in a vacuolar protease-deficient strain abolished this APP cleavage/release, and APP cleavage/release could be restored by introduction of MKC7 or YAP3 on a single copy plasmid. Purified Mkc7p cleaved an internally quenched fluorogenic APP peptide substrate at the ␣-secretase cleavage site. Measurement of proteolytic activity either in yeast homogenates or on the yeast cell surface revealed that most Mkc7p and Yap3p activities were localized at the cell surface. These results establish a molecular basis for ␣-secretase-type cleavage in yeast and support the generally held concept that ␣-secretase cleavage of APP occurs at the cell surface.The ␤-amyloid precursor protein (APP), 1 a type-1 transmembrane protein, can undergo at least two alternative pathways of proteolytic processing. ␤-Amyloid peptide (A␤), the component of cerebral plaques associated with Alzheimer's disease, is produced by proteolytic processing of APP and includes 28 residues of the N-terminal lumenal domain and the first 12-14 residues of the transmembrane domain (1). Alternatively, cleavage of a Lys-Leu bond the lumenal portion of the A␤ region of APP by a membrane protein-solubilizing proteolytic activity, termed "␣-secretase," releases the lumenal portion of APP as a soluble protein and precludes the formation of A␤ (1).There has been intensive interest in identifying the enzymes that cleave APP to determine their relationship to the etiology of Alzheimer's disease. However, the responsible molecules remain unknown. In particular, molecular identification of ␣-secretase could provide insight into the general mechanism of integral membrane protein cleavage (2) and clarify the role of this enzyme in the pathogenesis of Alzheimer's disease. We have reported that the yeast Saccharomyces cerevisiae has ␣-secretase-like activities: human APP expressed in S. cerevisiae undergoes proteolytic cleavage at the ␣-secretase site (3, 4). Existence of an ␣-secretase-like activity was also observed in insect cells (5), suggesting that the enzymes are widely conserved among metazoans. Therefore, identification of yeast secretases could lead to the discovery of counterparts among other phyla. Here we demonstrate that homologous glycosyl-phosphatidylinositol (GPI)-linked aspartyl proteases, Mkc7p (6) and Yap3p (7, 10 -12), termed "yapsin" (33), are responsible for ␣-secretase cleavage of human APP in yeast. EXPERIMENTAL PROCEDURESYeast Media and Strains-Liquid synthetic media were as described (13). All strains were derived from the W303 background as described (14). The str...

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“…The nonsecretory, amyloidogenic pathway of /IAPP metabolism, mediated by unknown proteases such as /I-secretase in an intracellular acidic compartment, accounts for '-~70%of total /IAPP metabolism; a key processing intermediary is A/I-peptide-containing CT fragments, which may be further processed to A/I in cultured cells Gandy et al, 1992a,b;Knops et al, 1992;Shoji et al, 1992;Cheung et al, 1994) and in human brains (Golde etal., 1992;Kametani et al, 1994). Some of the enzymes suggested by various groups as /I-secretase candidates include cathepsin U (Sahasrabudhe et al, 1993;Brown et al, 1996), calcium-activated serine protease (Abraham et al, 1991), multicatalytic protease (Ishiura et al., 1989), chymase or mast cell protease (Nelson et al, 1993), clipsin (Nelson and Siman, 1990), calpain I (Siman et al, 1990, the metalloprotease protease EC 3.4.24.15 (McDermott et al, 1992;Papastoitsis et al, 1994), and cathepsin D (Ladror et al, 1994;Brown et al, 1996).…”

Section: Generation Of Amyloidogenic Ct Fragmentsmentioning

confidence: 99%

An Etiological Role of Amyloidogenic Carboxyl‐Terminal Fragments of the β‐Amyloid Precursor Protein in Alzheimer's Disease

Suh

1997

Journal of Neurochemistry

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Amyloid~3protein (A/3), 39-43 amino acids long, is the principal constituent of the extracellular amybid deposits in brain that are characteristic of Alzheimer's disease (AD). Several lines of evidence indicate that A~3 may play an important role in the pathogenesis of AD. However, there are several discrepancies between the production of A/3 and the development of the disease. Thus, A/may not be the sole active fragment of~3-amybid precursor protein (~3APP) in the neurotoxicity associated with AD. Consequently, the possible effects of other cleaved products of /3APP need to be explored. The recent concentration on other potentially amyloidogenic products of /IAPP has produced interesting candidates, the most promising of which are the amyloidogenic carboxyl-terminal (CT) fragments of~3APP. This review discusses a possible etiological role of CT fragments of /3APP in AD. Key Words: Amyloid precursor proteinCarboxyl-terminal peptide-Amyloid /3 protein -Etiological role-Alzheimer's disease. J. Neurochem. 68, 1781Neurochem. 68, -1791Neurochem. 68, (1997.Alzheimer' s disease (AD) is characterized by amybid deposits in senile plaques, in neurofibrillary tangles, and on the walls of cerebral blood vessels and dystrophic neurites, gliosis, and loss of neuronal synapses. various evidence indicates that amyloid~3pro-tein (A/3), which is a principal constituent of senile plaques (Glenner and Wong, 1984), may play an important role in the pathogenesis of AD (for review, see Selkoe, 1994;Checler, 1995). The A/I is composed of 39-43 amino acids and proteolytically derived from larger /3-amyloid precursor protein (/3APP) isoforms of 695, 714, 751, and 770 amino acids (Kang et al., 1987). Several mutations of /IAPP around the A/I domain have been discovered in certain types of earlyonset familial AD (FAD) (for review, see Tanzi et al., 1993), supporting its causal role in the pathogenesis of AD.To date, at least three processing pathways have been described: the nonamyloidogenic secretory pathway (a-secretase), which releases soluble ectodomain and prevents A/I formation (Esch et al., 1990); the endosomal-lysosomal pathway, in which some cell surface /IAPPs are reinternalized (Haass et al., 1 992a,b) and cleaved around at the N-terminus of the A/I sequence by /3-secretase to produce A/3-bearing amyloidogenic breakdown products of various sizes (14-22 kDa) Golde et al., 1992; Haass et al., 1992a,b;Shoji et al., 1992;Tamaoka et al., 1992) and subsequently possibly cleaved by 'ysecretase to release soluble 4-kDa A/I (Koo and Squazzo, 1994); and the 4-kDa A/I-producing pathway (/3-and y-secretases), involving coated pit-mediated endocytosis, which may be independent of the constitutive secretory pathway (Koo and Squazzo, 1994).Classically, A/I is the marker for AD, and it has been linked to the accompanying neurodegeneration (see, e.g., Sisodia et al., 1990). Several lines of evidence suggest that the overexpression of /3APP and the subsequent production of A/I could be linked to the genesis of AD (for review, see ...

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Characterization by radiosequencing of the carboxyl-terminal derivatives produced from normal and mutant amyloid β protein precursors

Cited by 25 publications

References 25 publications

Secretory cleavage site of Alzheimer amyloid precursor protein is heterogeneous in Down's syndrome brain

Secretory cleavage site of Alzheimer amyloid precursor protein is heterogeneous in Down's syndrome brain

Involvement of Cell Surface Glycosyl-phosphatidylinositol-linked Aspartyl Proteases in α-Secretase-type Cleavage and Ectodomain Solubilization of Human Alzheimer β-Amyloid Precursor Protein in Yeast

An Etiological Role of Amyloidogenic Carboxyl‐Terminal Fragments of the β‐Amyloid Precursor Protein in Alzheimer's Disease

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