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A validated antibody panel for the characterization of tau post-translational modifications
- Source :
- Molecular neurodegeneration 12(1), 87 (2017). doi:10.1186/s13024-017-0229-1, Molecular Neurodegeneration, Vol 12, Iss 1, Pp 1-19 (2017), Molecular Neurodegeneration
- Publication Year :
- 2017
- Publisher :
- Biomed Central, 2017.
-
Abstract
- Tau is a microtubule-binding protein, which is subject to various post-translational modifications (PTMs) including phosphorylation, methylation, acetylation, glycosylation, nitration, sumoylation and truncation. Aberrant PTMs such as hyperphosphorylation result in tau aggregation and the formation of neurofibrillary tangles, which are a hallmark of Alzheimer’s disease (AD). In order to study the importance of PTMs on tau function, antibodies raised against specific modification sites are widely used. However, quality control of these antibodies is lacking and their specificity for particular modifications is often unclear. In this study, we first designed an online tool called ‘TauPTM’, which enables the visualization of PTMs and their interactions on human tau. Using TauPTM, we next searched for commercially available antibodies against tau PTMs and characterized their specificity by peptide array, immunoblotting, electrochemiluminescence ELISA and immunofluorescence technologies. We demonstrate that commercially available antibodies can show a significant lack of specificity, and PTM-specific antibodies in particular often recognize non-modified versions of the protein. In addition, detection may be hindered by other PTMs in close vicinity, complicating the interpretation of results. Finally, we compiled a panel of specific antibodies and show that they are useful to detect PTM-modified endogenous tau in hiPSC-derived neurons and mouse brains. This study has created a platform to reliably and robustly detect changes in localization and abundance of post-translationally modified tau in health and disease. A web-based version of TauPTM is fully available at http://www.tauptm.org .
- Subjects :
- 0301 basic medicine
Glycosylation
SUMO protein
Hyperphosphorylation
Mice, Transgenic
tau Proteins
Computational biology
lcsh:Geriatrics
Immunofluorescence
Methylation
lcsh:RC346-429
03 medical and health sciences
Cellular and Molecular Neuroscience
chemistry.chemical_compound
Mice
0302 clinical medicine
Antibody Specificity
ddc:570
medicine
Animals
Humans
Phosphorylation
Molecular Biology
lcsh:Neurology. Diseases of the nervous system
biology
medicine.diagnostic_test
Antibody validation
Brain
Acetylation
metabolism [tau Proteins]
immunology [tau Proteins]
lcsh:RC952-954.6
030104 developmental biology
chemistry
metabolism [Brain]
biology.protein
Post-translational modification
Neurology (clinical)
Tau
Antibody
Alzheimer’s disease
Neuroscience
Protein Processing, Post-Translational
030217 neurology & neurosurgery
Research Article
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- Molecular neurodegeneration 12(1), 87 (2017). doi:10.1186/s13024-017-0229-1, Molecular Neurodegeneration, Vol 12, Iss 1, Pp 1-19 (2017), Molecular Neurodegeneration
- Accession number :
- edsair.doi.dedup.....e0ce594d4a959e8f7b7a349262390c18