Application of TurboID (DIA) Proteomics

Nara. C. Marella

Project Collaborators

• George Lin PhD
• Tuan-Ahn Nguyen PhD
• Stefan Kubicek PhD
• Molecular Discovery Platform of CeMM

Nuclear Metabolism & Gene Expression

• MTHFD1 is a folate metabolic pathway enzyme
• Catalyzes reactions that form substrates for methionine, thymidylate, and de novo purine syntheses
• BRD4 is regulator of chromatin structure and transcription
• Inhibition of either BRD4 or MTHFD1 causes similar changes in nuclear metabolite composition and gene expression

Proximity Labeling of Nuclear Metabolism Enzymes

• Biotin Ligase tagged to selected nucleotide metabolism enzymes such as NUDT5, PPAT etc.
• DMSO as negative control
• Treatment conditions include Adenosine, dNUDT5
• Either C or N Terminal tagging to observe active regions

Spectronaut

• Spectral Library from RAW files of combined (Control and Treatment) conditions
• Higher mass tolerance
• no default imputation
• default normalization
• PG Quanity export

Thank you!

Any questions?