Characterizing interactions of ER resident proteins in situ through the
YST-PPI method
Abstract
The mutual interactions of ER resident proteins in the ER maintain its
functions, prompting the protein folding, modification, and
transportation. Here, a new method, named YST-PPI (YESS-based Split fast
TEV protease System for Protein-protein Interaction) was developed,
targeting the characterization of protein interactions in ER. YST-PPI
method integrated the YESS system, split-TEV technology and endoplasmic
reticulum retention signal peptide (ERS) to provide an effective
strategy for studying ER in situ PPIs in a fast and quantitative manner.
The interactions among 15 ER resident proteins of S. cerevisiae were
explored using the YST-PPI system, and their interaction network map was
constructed, in which more than 74 interacting resident protein pairs
were identified. Our studies also showed that Lhs1p plays a critical
role in regulating the interactions of most of the ER resident proteins,
expect the Sil1p, indicating its potential role in controlling the ER
molecular chaperones. Moreover, the mutual interaction revealed by our
studies further confirmed that the ER resident proteins perform their
functions in a synergetic way and multimer complex might be formed
during the process.