Affinity purification coupled with mass spectrometry (AP–MS) and proximity-dependent biotinylation identification (BioID) methods have made substantial contributions to interaction proteomics studies. We have developed a GFP-tag AP-MS workflow to study the virus-host interactions of several medically and economically important viruses in Thailand. We are currently developing a proteomics platform for the dynamic visualization of signalosome landscape, where we combine the CRISPR-Cas9 genome editing with the BioID system. Cells will be engineered to produce the biotin ligase-tagged signaling protein of interest and BioID will be performed following the activation of the signaling pathway in a time-course manner. When paired with quantitative proteomics approach, the assay will be able to provide snapshots of both core and transient protein landscape of the signaling pathway over time.

1.Kovanich D, Saisawang C, Sittipaisankul P, Ramphan S, Kalpongnukul N, Somparn P,
Pisitkun T, Smith DR., 2019. Analysis of the Zika and Japanese Encephalitis Virus NS5 Interactomes. J Proteome Res, 18(8), 3203-3218.

The dataset of our flavivirus NS5 interacting proteins expands the current interaction landscape of NS5 and uncovers several NS5 targets that are new to flavivirus biology.