Applications of Amine-Reactive Tandem Mass Tags (TMT) in Human Neuroproteomics
Neuroproteomics is a complex field of life sciences due to the high complexity of the brain. This area comprises different pathophysiological conditions such as normal neurodevelopment, neurovascular disorders, and neurodegenerative disorders. A massive amount of studies have been performed using proteomics to increase the knowledge in this topic. However, there are still a lot more to explore. The most common proteomic techniques for investigating the different stages and conditions in neurodevelopment and diseases have mainly been based on two-dimensional gel electrophoresis (2-DE). More recently, the use of amine-reactive tandem mass tags (TMT) has also contributed to increase the understanding of the brain and associated disorders. The TMT can simultaneously compare up to ten samples and is compatible with a variety of biological samples. The proteins are labeled, pooled and co-eluted, and analyzed by LC-MS/MS. The multiplexing allows different designs and comparisons between the samples. Therefore the method is highly recommendable for, e.g., biomarker discovery in the neuroproteomic field. In this chapter the TMT 10-plex method will be detailed for use with three different brain proximal samples: cerebrospinal fluid (CSF), brain tissue, and neurons.
Key wordsMass spectrometry Isobaric tagging TMT Quantitative proteomics Neurodegenerative disorders Biomarker
- 10.Ye H, Boyne MT 2nd, Buhse LF, Hill J (2013) Direct approach for qualitative and quantitative characterization of glycoproteins using tandem mass tags and an LTQ Orbitrap XL electron transfer dissociation hybrid mass spectrometer. Anal Chem 85(3):1531–1539. doi: 10.1021/ac3026465 CrossRefPubMedGoogle Scholar
- 16.Licker V, Turck N, Kovari E, Burkhardt K, Cote M, Surini-Demiri M, Lobrinus JA, Sanchez JC, Burkhard PR (2014) Proteomic analysis of human substantia nigra identifies novel candidates involved in Parkinson’s disease pathogenesis. Proteomics 14(6):784–794. doi: 10.1002/pmic.201300342 CrossRefPubMedGoogle Scholar
- 24.Deutsch EW, Overall CM, Van Eyk JE, Baker MS, Paik YK, Weintraub ST, Lane L, Martens L, Vandenbrouck Y, Kusebauch U, Hancock WS, Hermjakob H, Aebersold R, Moritz RL, Omenn GS (2016) Human proteome project mass spectrometry data interpretation guidelines 2.1. J Proteome Res 15(11):3961–3970. doi: 10.1021/acs.jproteome.6b00392 CrossRefPubMedPubMedCentralGoogle Scholar