BMC Biotechnol 2017 Sep 2;17(1):69. Epub 2017 Sep 2.
Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai, 200438, China.
Download full-text PDF
Methods Mol Biol 2015 ;1346:151-68
Institute of Molecular Genetics and Cell Biology, Department of Biology, Ulm University, James-Franck-Ring N27, 89081, Ulm, Germany.
Protein interactions occur at certain times and at specific cellular places. The past years have seen a massive accumulation of binary protein-protein interaction data. The rapid increase of this context-free information necessitates robust methods to monitor protein interactions with temporal and spatial resolution in single cells. Read More
BMC Cell Biol 2013 Mar 11;14:14. Epub 2013 Mar 11.
Cambridge Institute for Medical Research, Hills Road, Cambridge CB2 2XY, UK.
Background: The split-ubiquitin system monitors interactions of transmembrane proteins in yeast. It is based on the formation of a quasi-native ubiquitin structure upon interaction of two proteins to which the N- and C-terminal halves of ubiquitin have been fused. In the system we use here ubiquitin formation leads to proteolytic cleavage liberating a transcription factor (PLV) from the C-ubiquitin (C) fusion protein which can then activate reporter genes. Read More
Mol Biol Cell 1999 Aug;10(8):2519-30
Max-Delbrück-Laboratorium, D-50829 Köln, Germany.
The split-Ubiquitin (split-Ub) technique was used to map the molecular environment of a membrane protein in vivo. Cub, the C-terminal half of Ub, was attached to Sec63p, and Nub, the N-terminal half of Ub, was attached to a selection of differently localized proteins of the yeast Saccharomyces cerevisiae. The efficiency of the Nub and Cub reassembly to the quasi-native Ub reflects the proximity between Sec63-Cub and the Nub-labeled proteins. Read More
Genome Res 2003 Jul;13(7):1744-53
Institute of Veterinary Biochemistry and Molecular Biology, University of Zurich-Irchel, CH-8057 Zurich, Switzerland.
Analysis of membrane protein interactions is difficult because of the hydrophobic nature of these proteins, which often renders conventional biochemical and genetic assays fruitless. This is a substantial problem because proteins that are integral or associated with membranes represent approximately one-third of all proteins in a typical eukaryotic cell. We have shown previously that the modified split-ubiquitin system can be used as a genetic assay for the in vivo detection of interactions between the two characterized yeast transmembrane proteins, Ost1p and Wbp1p. Read More