Strategies for examination of Alzheimer’s disease amyloid precursor protein isoforms

Newton, Jillian; Parkinson, David K.; Clench, Malcolm R.

doi:10.1007/s00216-006-0462-x

Cited by 9 publications

(6 citation statements)

References 36 publications

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“…APP is a transmembrane glycoprotein that contains 695, 751 or 770 amino acid residues depending upon the splice variance [22]. Synthesis of APP together with its N-terminal glycosylation occurs within the endoplasmic reticulum (ER), immature APP then moves to the Golgi apparatus for maturation and, thereafter, it eventually is transported towards the cell membrane [10].…”

mentioning

confidence: 99%

Memantine treatment decreases levels of secreted Alzheimer's amyloid precursor protein (APP) and amyloid beta (Aβ) peptide in the human neuroblastoma cells

Ray

Banerjee²,

Greig

et al. 2010

Neuroscience Letters

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Memantine, an uncompetitive NMDA receptor antagonist, is a FDA approved drug used for the treatment of moderate to severe Alzheimer’s disease (AD). Several studies have documented protective roles of memantine against amyloid beta peptide (Aβ)– mediated damage to neurons in both in vitro and in vivo models. Memantine is also effective in reducing amyloid burden in the brain of APP transgenic mice. Currently the role of memantine in the Aβ– mediated neurodegenerative cascade, including APP metabolism is being understood [1]. Herein, we investigated the effect of memantine on levels of the secreted form of Aβ precursor protein (APP), secreted Aβ and cell viability markers under short/acute conditions. We treated neuronal SK-N-SH cells with 10μM memantine and measured levels of secreted total APP (sAPP), APPα isoform and Aβ (1-40) in a time dependent manner for up to 24 hours. Memantine significantly decreased the levels of the secreted form of sAPP, sAPPα and Aβ (1-40) compared to vehicle- treated cells. This change started as early as 8 hour and continued for up to 24 hour of drug treatment. Unlike sAPP, a slight non-significant increase in total intracellular APP level was observed in 24-hour treated memantine cells. Taken together, these results suggest a role for memantine in the transport or trafficking of APP molecules away from the site of their proteolytic cleavage by the secretase enzymes. Such a novel property of memantine warrants further study to define its therapeutic utility.

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mentioning

confidence: 99%

Memantine treatment decreases levels of secreted Alzheimer's amyloid precursor protein (APP) and amyloid beta (Aβ) peptide in the human neuroblastoma cells

Ray

Banerjee²,

Greig

et al. 2010

Neuroscience Letters

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“…Alzheimer's disease Cell cultures: A central hallmark of Alzheimer's pathology is the neuritic plaques (composed primarily of Abeta proteins) present in brain tissue (Kidd, 1964;Roth et al, 1966). As distinct APP isoforms might be differentially involved in AD pathogenesis, Newton et al (2006) examined whether a MS platform was appropriate for the identification of the APP isoforms APP 695 , APP 751 , and APP 770 , in an in vitromodel (see Table 1). Three 2D-PAGE protein spots corresponding to these three isoforms were detected.…”

Section: Proteomic Studies In Psychiatric Disordersmentioning

confidence: 99%

Proteomic research in psychiatry

et al. 2010

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Psychiatric disorders such as Alzheimer's disease, schizophrenia and mood disorders are severe and disabling conditions of largely unknown origin and poorly understood pathophysiology. An accurate diagnosis and treatment of these disorders is often complicated by their aetiological and clinical heterogeneity. In recent years proteomic technologies based on mass spectrometry have been increasingly used, especially in the search for diagnostic and prognostic biomarkers in neuropsychiatric disorders. Proteomics enable an automated high-throughput protein determination revealing expression levels, post-translational modifications and complex protein-interaction networks. In contrast to other methods such as molecular genetics, proteomics provide the opportunity to determine modifications at the protein level thereby possibly being more closely related to pathophysiological processes underlying the clinical phenomenology of specific psychiatric conditions. In this article we review the theoretical background of proteomics and its most commonly utilized techniques. Furthermore the current impact of proteomic research on diverse psychiatric diseases, such as Alzheimer's disease, schizophrenia, mood and anxiety disorders, drug abuse and autism, is discussed. Proteomic methods are expected to gain crucial significance in psychiatric research and neuropharmacology over the coming decade.

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“…Although A  is the actual marker for AD, the detection of a smaller water soluble peptide A 1-16 will be a step forward in the development of a rapid and cost-effective bioanalytical tool for AD diagnostics. The immunoassay with specific monoclonal DE2 antibodies raised against A 1-16 (Newton and Parkinson, 2006) was utilized in this work in conjunction with the method of TIRE. In addition, TIRE dynamic spectra measurements were performed to study the kinetics of the immune reaction and subsequent evaluation of the affinity constant.…”

Section: Introductionmentioning

confidence: 99%

“…G418-resistant cells were expanded into DMEM medium supplemented with 10% FBS and selected for APP 770 secretion by immunoblotting with DE2. The technology of production of APP 770 was described in detail in (Newton and Parkinson, 2006) The substrates for TIRE measurements were prepared by consecutive thermal evaporation of chromium (31 nm) and gold (303 nm) layers on standard microscopic glass slides without breaking the vacuum of about 10 -6 Torr, using the Edwards E306A evaporation unit. The presence of thin Cr layer improves the adhesion of Au layer to glass.…”

Section: Introductionmentioning

confidence: 99%

Detection of β-amyloid peptide (1–16) and amyloid precursor protein (APP770) using spectroscopic ellipsometry and QCM techniques: A step forward towards Alzheimers disease diagnostics

Mustafa

Nabok

Parkinson

et al. 2010

Biosensors and Bioelectronics

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A highly sensitive method of spectroscopic ellipsometry in total internal reflection mode (TIRE) was exploited for detecting β-amyloid peptide (Aβ(1-16)) in the direct immune reaction with monoclonal DE2 antibodies (raised against Aβ(1-16)) electrostatically immobilised on the surface of gold. A rapid detection of Aβ(1-16) in a wide range of concentrations from 5 μg/ml down to 0.05 ng/ml was achieved using a cost-effective and label-free direct immunoassay format. TIRE dynamic spectral measurements proved that the immune reaction between DE2 monoclonal antibodies and Aβ(1-16) is highly specific with the affinity constant K(D)=1.46×10(-8) mol/l. The same DE2 antibodies were utilised for detection of amyloid precursor protein APP(770), a larger protein containing Aβ(1-16) domain, using the quartz crystal microbalance (QCM) measurements in liquid. A combination of QCM and TIRE kinetics results allowed the evaluation of the originally unknown concentration of APP(770) in complete medium solution containing other proteins, salts, and amino acids.

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Strategies for examination of Alzheimer’s disease amyloid precursor protein isoforms

Cited by 9 publications

References 36 publications

Memantine treatment decreases levels of secreted Alzheimer's amyloid precursor protein (APP) and amyloid beta (Aβ) peptide in the human neuroblastoma cells

Memantine treatment decreases levels of secreted Alzheimer's amyloid precursor protein (APP) and amyloid beta (Aβ) peptide in the human neuroblastoma cells

Proteomic research in psychiatry

Detection of β-amyloid peptide (1–16) and amyloid precursor protein (APP770) using spectroscopic ellipsometry and QCM techniques: A step forward towards Alzheimers disease diagnostics

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