title: Development of a membrane-anchored ligand and receptor yeast two-hybrid system for ligand-receptor interaction identification.
abstract: Identifying interactions between ligands and transmembrane receptors is crucial for understanding the endocrine system. However, the present approaches for this purpose are still not capable of high-throughput screening. In this report, a membrane-anchored ligand and receptor yeast two-hybrid (MALAR-Y2H) system was established. In the method, an extracellular ligand is linked with an intracellular split-ubiquitin reporter system via a chimeric transmembrane structure. Meanwhile, the prey proteins of transmembrane receptors are fused to the other half of the split-ubiquitin reporter system. The extracellular interaction of ligands and receptors can lead to the functional recovery of the ubiquitin reporter system in yeast, and eventually lead to the expression of report genes. Consequently, the system can be used to detect the interactions between extracellular ligands and their transmembrane receptors. To test the efficiency and universality of the method, interactions between several pairs of ligands and receptors of mouse were analyzed. The detecting results were shown to be thoroughly consistent with the present knowledge, indicating MALAR-Y2H can be utilized for such purpose with high precision, high efficiency and strong universality. The characteristics of the simple procedure and high-throughput potential make MALAR-Y2H a powerful platform to study protein-protein interaction networks between secreted proteins and transmembrane proteins.
authors.affiliation: Laboratory of Regeneromics, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Li
authors.affiliation: Laboratory of Regeneromics, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Gao
authors.affiliation: Laboratory of Regeneromics, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Han
authors.affiliation: Laboratory of Regeneromics, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Yinjie
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Shanghai Municipality Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Guan
authors.affiliation: Shanghai Municipality Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Shanghai Municipality Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Laboratory of Regeneromics, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Han
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D008024:Ligands; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D020798:Two-Hybrid System Techniques
chemical_list: D008024:Ligands; D008565:Membrane Proteins