Affiliation:
1. State Key Laboratory of Biocatalysis and Enzyme Engineering Hubei Collaborative Innovation Center for Green Transformation of Bio‐resources Hubei Key Laboratory of Industrial Biotechnology School of Life Sciences Hubei University Wuhan China
2. College of Life Science and Technology Beijing University of Chemical Technology Beijing China
Abstract
AbstractThe mutual interactions of endoplasmic reticulum (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, most being identified molecular chaperones, 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, except 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 cooperative way and a multimer complex might be formed during the process.
Funder
National Key Research and Development Program of China
National Natural Science Foundation of China
Fundamental Research Funds for the Central Universities