Snapshot Analysis of Protein Complexes Using Proteomic Technology

抄録

Proteomic technologies allow a comprehensive study of multi-protein complexes that carry out many cellular functions in a higher-order network in the cell. The use of tagged proteins as affinity bait, coupled with mass spectrometric identification, enables us to isolate almost any functional protein complex or its synthetic intermediates that might represent snapshots of nascent functional protein complexes at particular stages of its biogenesis and to identify their constituents-some of which show dynamic changes for association with the intermediates at various stages of the biogenesis. The idea behind this snapshot analysis is that some of the associated proteins in one initially isolated complex can also be present in other precursor complexes, and thus would allow the purification of intermediates from different stages of biogenesis. Initially, we had applied this approach to the analysis of protein constituents of pre-ribosomes, and is now expanded this to the analysis of not only protein but also RNA constituents of small nuclear ribonucleoprotein (snRNP) intermediates formed during spliceosome biogenesis in cooperation with Dr. Isobe of Tokyo Metropolitan University. In this presentation, I would like to talk about mainly current status of the snapshot analysis of snRNP complexes.