Protein-protein interactions as they appear in functional protein complexes have been studied in high throughput fashion in yeast, through yeast 2-hybrid on the one hand and affinity purification followed by mass spectrometry (AP-MS) on the other hand. We have developed a unique AP-MS platform for protein complex isolation from plant cells.
Complex purifications are performed through tandem affinity purification (TAP) from Arabidopsis cell suspension cultures or whole seedlings. Co-purified proteins are identified by mass spectrometry in close collaboration with the Proteomics Core Facility at VIB. We have been able to isolate the T-plate complex driving endocytosis in plants, and the AN3 complex driving cell proliferation in leaf. Besides, our publications holds numerous examples of successful protein complex purifications for proteins present in diverse cellular pathways, in different organelles or the cytosol, both soluble or membrane associated. More recently, our attention has also moved towards the optimization of tools to isolate transient or very temporal protein-protein interactions.