Differential Epitope Identification of Antibodies Against Intracellular Domains of Alzheimer's Amyloid Precursor Protein Using High Resolution Affinity-mass Spectrometry

2007 ◽  
pp. 339-354 ◽  
Author(s):  
Xiaodan Tian ◽  
Madalina Maftei ◽  
Markus Kohlmann ◽  
Bernadette Allinquant ◽  
Michael Przybylski
Keyword(s):  
Mass Spectrometry ◽  
High Resolution ◽  
Amyloid Precursor Protein ◽  
Precursor Protein ◽  
Epitope Identification ◽  
Intracellular Domains

Identification and Structural Characterisation of Carboxy-Terminal Polypeptides and Antibody Epitopes of Alzheimer's Amyloid Precursor Protein Using High-Resolution Mass Spectrometry

2005 ◽  
Vol 11 (5) ◽  
pp. 547-555 ◽  
Author(s):  
Xiaodan Tian ◽  
Roxana Cecal ◽  
JoAnne McLaurin ◽  
Marilena Manea ◽  
Raluca Stefanescu ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Resolution ◽  
Amyloid Precursor Protein ◽  
Vaccine Development ◽  
Precursor Protein ◽  
Therapeutic Effects ◽  
Amyloid A ◽  
Carboxy Terminal ◽  
Ft Icr Ms ◽  
Ft Icr

Alzheimer's disease (AD) is the most common cause for human age-related dementia, characterised by formation of diffuse plaques in the brain that are directly involved in AD pathogenesis. The major component of AD plaques is ß-amyloid, a 40 to 42 amino acid polypeptide derived from the amyloid precursor protein (APP) by proteolytic degradation involving the specific proteases, ß-and γ-secretase acting at the N- and C-terminal cleavage site, respectively. In this study, we have prepared polypeptides comprising the carboxy-terminal and transmembrane sequences of APP, by bacterial expression and chemical synthesis, as substrates for studying the C-terminal processing of APP and its interaction with the γ-secretase complex. Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) was used as a major tool for structure analysis. Immunisation of transgenic mouse models of AD with Aß42 has been recently shown to be effective to inhibit and disaggregate Aß-fibrils, and to reduce AD-related neuropathology and memory impairments. However, the mechanism underlying these therapeutic effects as yet has been unclear. Using proteolytic epitope excision from immune complexes in combination with FT-ICR-MS, we identified the epitope recognised by the therapeutically active antibody as the N-terminal Aß(4–10) sequence; this soluble, nontoxic epitope opens new lead structures for AD vaccine development. A monoclonal antibody (Jonas; JmAb) directed against the cytosolic APP domain was used in studies of APP biochemistry and metabolism. Here, we report the identification of the epitope recognised by the JmAb, using the combination of epitope excision and peptide mapping by FT-ICR-MS. The epitope was determined to be located at the C-terminal APP(740–747) sequence; it was confirmed by ELISA binding assays and authentic synthetic peptides and will be an efficient tool in the development of new specific vaccines. These results demonstrate high-resolution FT-ICR-MS as a powerful method for characterising biochemical pathways and molecular recognition structures of APP.

scholarly journals Co-localization of the amyloid precursor protein and Notch intracellular domains in nuclear transcription factories

2010 ◽  
Vol 31 (1) ◽  
pp. 58-73 ◽  
Author(s):  
Uwe Konietzko ◽  
Zoë V. Goodger ◽  
Michelle Meyer ◽  
Bernhard M. Kohli ◽  
Jérôme Bosset ◽  
...  
Keyword(s):  
Amyloid Precursor Protein ◽  
Precursor Protein ◽  
Transcription Factories ◽  
Intracellular Domains

scholarly journals Amyloid Precursor Protein and Notch Intracellular Domains are Generated after Transport of their Precursors to the Cell Surface

Traffic ◽  
2006 ◽  
Vol 7 (4) ◽  
pp. 408-415 ◽  
Author(s):  
Christoph Kaether ◽  
Stephanie Schmitt ◽  
Michael Willem ◽  
Christian Haass
Keyword(s):  
Cell Surface ◽  
Amyloid Precursor Protein ◽  
Precursor Protein ◽  
Intracellular Domains

An on-flow assay for screening of β-secretase ligands by immobilised capillary reactor-mass spectrometry

2017 ◽  
Vol 9 (14) ◽  
pp. 2189-2196 ◽  
Author(s):  
Adriana Ferreira Lopes Vilela ◽  
Carmen Lúcia Cardoso
Keyword(s):  
Mass Spectrometry ◽  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Amyloid Precursor Protein ◽  
Mass Spectrometer ◽  
Precursor Protein ◽  
Enzyme Reactor ◽  
Β Amyloid

The enzyme β-secretase1 (BACE1) initiates the cleavage of the Ab amyloid precursor protein (APP), to generate and aggregate β-amyloid (Ab) peptides, which are implicated in the pathogenesis of Alzheimer's disease (AD). Herein, a BACE1 immobilised capillary enzyme reactor (ICER) attached to a mass spectrometer for the on-flow screening of ligands was prepared.

scholarly journals Tandem Mass Spectrometry as Strategy for the Selective Identification and Quantification of the Amyloid Precursor Protein Tyr682 Residue Phosphorylation Status in Human Blood Mononuclear Cells

Biomolecules ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 1297
Author(s):  
Pierluigi Reveglia ◽  
Rosarita Nasso ◽  
Antonella Angiolillo ◽  
Lucia Lecce ◽  
Carmela Paolillo ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Amyloid Precursor Protein ◽  
Mononuclear Cells ◽  
Clinical Signs ◽  
Precursor Protein ◽  
Tandem Mass ◽  
Blood Mononuclear Cells ◽  
Phosphorylated Peptide ◽  
Identification And Quantification

Background: Alzheimer’s disease (AD) is a devastating neurodegenerative disease without guidelines for early diagnosis or personalized treatment. Previous studies have highlighted a crucial role of increasing phosphorylation levels of the amyloid precursor protein (APP) Tyr682 residue in predicting neuronal deficits in AD patients. However, the lack of a method for the identification and quantification of Tyr682 phosphorylation levels prevents its potential clinical applications. Methods: Here we report a method to identify and quantify APP Tyr682 phosphorylation levels in blood mononuclear cells of AD patients by tandem mass spectrometry (tMS). Results: This method showed excellent sensitivity with detection and quantification limits set respectively at 0.035 and 0.082 ng injected for the phosphorylated peptide and at 0.02 and 0.215 ng injected for the non-phosphorylated peptide. The average levels of both peptides were quantified in transfected HELA cells (2.48 and 3.53 ng/μg of protein, respectively). Preliminary data on 3 AD patients showed quantifiable levels of phosphorylated peptide (0.10–0.15 ng/μg of protein) and below the LOQ level of non-phosphorylated peptide (0.13 ng/μg of protein). Conclusion: This method could allow the identification of patients with increased APP Tyr682 phosphorylation and allow early characterization of molecular changes prior to the appearance of clinical signs.

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