Yeast-two hybrid systems have commonly been used to probe protein-protein interactions. However, this requires the creation of recombinant proteins, which may alter protein binding. One chemical-biology-based alternative is crosslinking, using a synthetic amino acid encoded into one of the proteins. The aliphatic diazirine amino acid, 3'- azibutyl-N-carbamoyl-lysine (Abk) has previously been incorporated into protein by using the wild-type M.barkeri pyrrolsysl tRNA/tRNA synthetase pair. UV light initiates the formation of a covalent bond between the two proteins. Schultz et al report that a modified aminoacyl-tRNA synthetase in combination with pyrrolysyl tRNA can significantly reduce a major problem with this system in mammalian cells- low coding efficiency.
"Ye, Z., Desai, H., Bair, M., Williams, G.J. (2011) A novel photocrosslinking assay for reporting protein interactions in polyketide and fatty acid synthases. Mol. BioSys, In press."
ReplyDeleteThis paper from Zhixia uses the same approach to probe protein-protein interactions but with a different photo-crosslinking unnatural amino acid.
Peter Schultz bothering to improve this assay for studying protein-protein interactions in mammalian cells just shows how useful such an assay is.
Well, we actually scooped the Schultz lab this time - certainly does not happen often: http://pubs.rsc.org/en/Content/ArticleLanding/2011/SC/C0SC00373E
ReplyDelete:)