Abstract
Here, we describe a capillary electrophoresis method for evaluating proteolysis of the amyloid beta peptide (Aβ) 1–40. This method is suitable for kinetic studies, demands little specialized equipment, and consumes only small quantities of a commercially available substrate whose physiological accumulation is thought to underlie the development of Alzheimer’s disease.
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References
Glenner GG, Wong CW (1984) Alzheimer’s disease: initial report of the purification and characterization of a novel cerebrovascular amyloid protein. Biochem Biophys Res Comm 120:885–890
Hardy J, Selkoe DJ (2002) The amyloid hypothesis of Alzheimer’s disease: progress and problems on the road to therapeutics. Science 297:353–356
McDermott JR, Gibson AM (1997) Degradation of Alzheimer’s beta-amyloid protein in human and rat brain peptidases: involvement of insulin-degrading enzyme. Neurochem Res 22:49–56
Vekrellis K, Ye Z, Qiu WQ, Walsh D, Hartley D, Chesneau V et al (2000) Neurons regulate extracellular levels of amyloid-beta protein via proteolysis by insulin-degrading enzyme. J Neurosci 20:1657–1665
Mukherjee A, Song E, Kihiko-Ehmann M, Goodman JP, St Pyrek J, Estus S et al (2000) Insulysin hydrolyzes amyloid-beta peptides to products that are neither neurotoxic nor deposit on amyloid plaques. J Neurosci 20:8745–8749
Miller BC, Eckman EA, Sambamurti K, Dobbs N, Chow KM, Eckman CB et al (2003) Amyloid beta peptide levels in brain are inversely correlated with insulysin activity levels in vivo. Proc Natl Acad Sci USA 100:6221–6226
Leissring MA, Farris W, Chang AY, Walsh DM, Wu X, Sun X et al (2003) Enhanced proteolysis of beta-amyloid in APP transgenic mice prevents plaque formation, secondary pathology, and premature death. Neuron 40:1087–1093
Varesio E, Rudaz S, Krause K-H, Veuthey J-L (2002) Nanoscale liquid chromatography and capillary electrophoresis coupled to electrospray mass spectrometry for the detection of amyloid beta peptide related to Alzheimer’s disease. J Chr A 974:135–142
Sabella S, Quaglia M, Lanni C, Racchi M, Govoni S, Caccialanza G et al (2004) Capillary electrophoresis studies on the aggregation process of beta-amyloid 1–42 and 1–40 peptides. Electrophoresis 25:3186–3194
Verpillot R, Otto M, Klafki H, Taverna M (2008) Simultaneous analysis by capillary electrophoresis of five amyloid peptides as potential biomarkers of Alzheimer’s disease. J Chr A 1214:157–164
Alper BJ, Schmidt WK (2009) A capillary electrophoresis method for evaluation of Aβ proteolysis in vitro. J Neurosci Methods 178:40–45
Gomori G (1955) Preparation of buffers for use in enzyme studies. Methods Enzymol 1:138–146
Alper BJ, Rowse JW, Schmidt WK (2009) Yeast Ste23p shares functional similarities with mammalian insulin-degrading enzymes. Yeast 11:595–610
Acknowledgment
We are grateful to Drs. Nandu Menon, Alan Przybyla, and Zachary Wood (all of the University of Georgia, Department of Biochemistry and Molecular Biology) for advice and assistance, and for access to experimental equipment and materials. Aβ 1–40 consumed during method development and analysis was a gift of rPeptide, Inc. (Athens, GA, USA).
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Alper, B.J., Schmidt, W.K. (2013). Evaluating Amyloid Beta (Aβ) 1-40 Degradation by Capillary Electrophoresis. In: Volpi, N., Maccari, F. (eds) Capillary Electrophoresis of Biomolecules. Methods in Molecular Biology, vol 984. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-296-4_19
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DOI: https://doi.org/10.1007/978-1-62703-296-4_19
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-295-7
Online ISBN: 978-1-62703-296-4
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