Epitope map of two polyclonal antibodies that recognize amyloid lesions in patients with Alzheimer's disease

Ghiso, Jorge; Wısnıewskı, Thomas; Vidal, Rubén; Rostagno, Agueda; Frangione, Blas

doi:10.1042/bj2820517

Cited by 11 publications

(14 citation statements)

References 37 publications

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“…4A, lanes 3 and 4), suggesting that the C-terminus of the 4G8-reactive APP may extend to Aj3 Lys2' or beyond. In agreement with recently published reports (Ghiso et al, 1992), SP28 antiserum failed to react with full-length APP from PC12 cells (data not shown). To examine whether human cells also have secretase I1 activity, we probed purified secreted APP isolated from human embryonic kidney cells (293 cells) transfected with APP,,, (Esch et al, 1990).…”

Section: Resultssupporting

confidence: 93%

“…These residues correspond to APP,,, amino acids 673-680. In agreement with recently published reports (Ghiso et al, 1992), SP28 antiserum failed to react with full-length APP from PC12 cells (data not shown). This antiserum specifically recognized the APP fraction retained in the 4G8 affinity column (Fig.…”

Section: J P Anderson Et Alsupporting

confidence: 93%

“…These residues correspond to APP,,, amino acids 673-680. SP28 does not recognize full-length APP or its major secreted form (Ghiso et al, 1992). This antiserum specifically recognized the APP fraction retained in the 4G8 affinity column (Fig.…”

Section: Resultsmentioning

confidence: 95%

“…Recently an antiserum (SP28) has been prepared that recognizes Aj3 AlaZ1-Lys2' (Fig. 1) (Ghiso et al, 1992). These residues correspond to APP,,, amino acids 673-680.…”

Section: Resultsmentioning

confidence: 99%

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An Alternative Secretase Cleavage Produces Soluble Alzheimer Amyloid Precursor Protein Containing a Potentially Amyloidogenic Sequence

Anderson

Chen

Kim³

et al. 1992

Journal of Neurochemistry

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Cell culture studies have shown that the Alzheimer amyloid precursor protein (APP) is secreted after full‐length APP is cleaved by a putative secretase at the Lys16‐Leu17 bond (secretase cleavage I) of the amyloid peptide sequence. Because this cleavage event is incompatible with amyloid production, it has been assumed that secreted APP cannot serve as a precursor of the amyloid depositions observed in Alzheimer's disease. Here we show that in neuronally differentiated PC12 cells and human kidney 293 cell cultures a portion of the secreted extracytoplasmic APP reacted specifically with both a monoclonal antibody recognizing amyloid protein residues Leu17‐Val24 and a polyclonal antiserum directed against amyloid protein residues Ala21‐Lys28. Furthermore, this APP failed to react with antisera recognizing the cytoplasmic domain of the full‐length protein. These data indicate the presence of an alternative APP secretase cleavage site (secretase cleavage II), C‐terminal to the predominant secretase cleavage I. Depending on the exact location of cleavage site II, potentially amyloidogenic secreted APP species may be produced.

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Section: Resultssupporting

confidence: 93%

Section: J P Anderson Et Alsupporting

confidence: 93%

Section: Resultsmentioning

confidence: 95%

“…Recently an antiserum (SP28) has been prepared that recognizes Aj3 AlaZ1-Lys2' (Fig. 1) (Ghiso et al, 1992). These residues correspond to APP,,, amino acids 673-680.…”

Section: Resultsmentioning

confidence: 99%

See 2 more Smart Citations

An Alternative Secretase Cleavage Produces Soluble Alzheimer Amyloid Precursor Protein Containing a Potentially Amyloidogenic Sequence

Anderson

Chen

Kim³

et al. 1992

Journal of Neurochemistry

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“…Monoclonal antibodies 4G8 (18) and 6E10 (37) and polyclonal antibodies SP40 (19), SGY1234 (20), and R1280 (21) have been described.…”

Section: Methodsmentioning

confidence: 99%

Protein phosphorylation inhibits production of Alzheimer amyloid beta/A4 peptide.

Buxbaum

Koo

Greengard

1993

Proc. Natl. Acad. Sci. U.S.A.

267

166

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The major component of amyloid plaque cores and cerebrovascular amylold deposits found in Alzheimer disease Is the P/A4 peptide, which is derived from the AIzheimer amylold protein precursor (APP). Recent evidence sugests that abnormalities in f/A4 peptide production or ,B/A4 peptide agregation may underlie cerebral amyloidosis.In the present study, treatment of cells with phorbol dibutyrate, which activates protein kinase C, and/or okadaic acid, which inhibits protein phosphatases 1 and 2A, reduced P/A4 peptide production by 50-80%. These effects were observed with APP6. and APP75s expressed in stablY btrasfected CHO cells, as well as with endogenous APP in human glioma (Hs 683) cells. Phorbol dibutyrate also decreased P/A4 peptide production in cells expressing various mutant forms of APP associated with familial Alzheimer disease, one of which was reported to m est greatly incrased P/A4 peptide production in cultured cells. Mastoparan and mastoparan X, compounds which can activate phospholipase C and hence protein khin C, also decreased f/A4 peptide production in CHO cells stably transfected with APP695. A model is presented in which decreases in 3/A4 peptide production can be achieved by accelerating the metabolism of APP through a nonamyloidgenic secretory pathway. Pulse-chase labeling of cells was carried out on confluent cell monolayers in six-well culture dishes (Corning) with 1 ml of methionine-free Dulbecco's modified Eagle's medium (DMEM) supplemented with 1 mCi (37 MBq) of [35S]methionine (EXPRE35S35S, NEN). Metabolic labeling was carried out for 2 hr, followed by a chase period of2 hr. The chase was initiated by replacing the labeling medium with DMEM containing excess unlabeled methionine. Two minutes after the start of the chase, the indicated test compounds were added. This protocol maximized the probability that any observed effects were attributable to changes in APP metabolism rather than APP transcription. After incubation, conditioned medium was centrifuged for 5 min at 10,000 x g to remove cellular debris. 3/A4 peptide and p3 were immunoprecipitated with an anti-p/A4 antibody (4G8 except where indicated), and secreted APP (APPs) was immunoprecipitated with an anti-APP NH2-terminal antibody (22C11). Immunoprecipitated APP fragments were subjected to SDS/ PAGE in either 10-20% gels with Tris/tricine buffer (for f/A4 peptide and p3) or 6% gels with Tris/glycine buffer (for APPs), autoradiographed, and quantified with a PhosphorImager (Molecular Dynamics). Analysis of variance followed by Fisher's post-hoc analysis was used to determine the significance of observed differences. RESULTSWhen the medium of metabolically labeled CHO cells stably transfected with cDNA encoding the 695-amino-acid isoform of APP (APP695) was subjected to immunoprecipitation with an antibody raised against a synthetic peptide corresponding to ,/A4 peptide, two small peptide bands of apparent molecular mass 3 and 2 kDa were observed (Fig. 1). These peptides had migration rates similar to those of two peptides...

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Complete cerebral ischemia with short-term survival in rats induced by cardiac arrest. I. Extracellular accumulation of Alzheimer's β-amyloid protein precursor in the brain

et al. 1994

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Epitope map of two polyclonal antibodies that recognize amyloid lesions in patients with Alzheimer's disease

Cited by 11 publications

References 37 publications

An Alternative Secretase Cleavage Produces Soluble Alzheimer Amyloid Precursor Protein Containing a Potentially Amyloidogenic Sequence

An Alternative Secretase Cleavage Produces Soluble Alzheimer Amyloid Precursor Protein Containing a Potentially Amyloidogenic Sequence

Protein phosphorylation inhibits production of Alzheimer amyloid beta/A4 peptide.

Complete cerebral ischemia with short-term survival in rats induced by cardiac arrest. I. Extracellular accumulation of Alzheimer's β-amyloid protein precursor in the brain

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