Abstract
Alzheimer’s disease (AD) is characterized by an early, asymptomatic phase (AsymAD) in which individuals exhibit amyloid-beta (Aβ) plaque accumulation in the absence of clinically detectable cognitive decline. Here we report an unbiased multiplex quantitative proteomic and phosphoproteomic analysis using tandem mass tag (TMT) isobaric labeling of human post-mortem cortex (n = 27) across pathology-free controls, AsymAD and symptomatic AD individuals. With off-line high-pH fractionation and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) on an Orbitrap Lumos mass spectrometer, we identified 11,378 protein groups across three TMT 11-plex batches. Immobilized metal affinity chromatography (IMAC) was used to enrich for phosphopeptides from the same TMT-labeled cases and 51,736 phosphopeptides were identified. Of these, 48,992 were quantified by TMT reporter ions representing 33,652 unique phosphosites. Two reference standards in each TMT 11-plex were included to assess intra- and inter-batch variance at the protein and peptide level. This comprehensive human brain proteome and phosphoproteome dataset will serve as a valuable resource for the identification of biochemical, cellular and signaling pathways altered during AD progression.
Highlights
Background & SummaryAlzheimer’s disease (AD) is the most common age-related neurodegenerative disease, currently affecting more than 40 million people worldwide[1,2]
AD is characterized by an early, asymptomatic phase (AsymAD) in which individuals exhibit amyloid-beta (Aβ) plaque accumulation in the absence of significant tau neurofibrillary tangles (NFT) and cognitive decline[3,4]
Percolator was used to filter peptide spectral matches (PSM) and peptides to a false discovery rate (FDR) of less than 1% using target-decoy strategy
Summary
Background & SummaryAlzheimer’s disease (AD) is the most common age-related neurodegenerative disease, currently affecting more than 40 million people worldwide[1,2]. 27 samples from 3 groups (n = 10 control, n = 8 AsymAD, and n = 9 AD) were used for brain proteome and phosphoproteome analyses. For the total proteome runs, a total of 164,034 unique peptides were identified that mapped to 11,378 protein groups at a 1% FDR on the peptide spectrum match (PSM) level across all batches, which represented 10,373 coding gene products.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
