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Proteomics Data and Methods

This page provides information about the data and methods used to generate the Proteomics evidence presented in Agora.

The Proteomics evidence for a specific gene can be viewed on the Evidence/Protein tab of that gene’s page in Agora. You can navigate to a gene’s page using search, from the Nominated Targets list, or from the Gene Comparison Tool. You can also compare Proteomics evidence across multiple genes using Agora’s Gene Comparison Tool.

If you have questions, suggestions, or feedback about Agora, please let us know here.

Data and Methods

Protein abundance has been quantified using two methods; liquid-free quantification (LFQ) and tandem mass tagged (TMT) spectrometry:

LFQ Data and Methods

LFQ data was generated from post-mortem brains of more than 500 individuals. Samples were taken from four human cohort studies, representing four different brain regions:

Cohort Study

Brain Region(s)

Banner Sun Health Research Institute (Banner)

Dorsolateral Prefrontal Cortex (DLPFC)

Baltimore Longitudinal Study on Aging (BLSA)

Middle Frontal Gyrus (MFG)

Mayo RNAseq (MAYO)

Temporal Cortex (TCX)

Mount Sinai Brain Bank (MSBB)

Anterior Prefrontal Cortex (AntPFC)

Samples were analyzed using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Protein abundance was quantified using liquid-free quantification (LFQ). Note that not all proteins are detected in all brain regions; for these proteins, the plots will show fewer than four brain regions.

Samples were harmonized within study by batch correction with COMBAT and regressing out Age, Sex, and Post Mortem Interval (PMI). Differential expression of AD versus controls was determined via ANOVA with an FDR of 0.05.

The processed data is available in Synapse through this link: LFQ Proteomics for Agora.

TMT Data and Methods

TMT data was generated from post-mortem brains of 400 individuals from the ROSMAP cohort:

Cohort Study

Brain Region

The Religious Orders Study and Memory and Aging Project (ROSMAP)

Dorsolateral Prefrontal Cortex (DLPFC)

Samples were analyzed using isobaric tandem mass tag (TMT) peptide labeling as described in the Round 1 Methods section of Proteomics (TMT quantitation). Batch correction, relative abundance calculation, and log2 transformation was performed as described here.

Differential expression of AD versus controls was determined using linear regression, adjusted for sex and post-mortem interval. Output consists of log2 fold change for AD cases versus controls (with controls as the reference group), 95% confidence intervals, and FDR-adjusted p-values.

The processed data is available in Synapse through this link: TMT Proteomics for Agora.

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