Genetically Directed Production of Recombinant, Isosteric and Nonhydrolysable Ubiquitin Conjugates.
作者： Mathew Stanley , Satpal Virdee
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摘要: We describe the genetically directed incorporation of aminooxy functionality into recombinant proteins by using a mutant Methanosarcina barkeri pyrrolysyl-tRNA synthetase/tRNACUA pair. This allows general production nonhydrolysable ubiquitin conjugates origin bioorthogonal oxime ligation. was exemplified preparation versions diubiquitin, polymeric chains and ubiquitylated SUMO. The exhibited unrivalled isostery with native isopeptide bond, as inferred from structural biophysical characterisation. Furthermore, functioned nanomolar inhibitors deubiquitylating enzymes were recognised linkage-specific antibodies. technology should provide versatile platform for development powerful tools studying elucidating cellular roles diverse polyubiquitin linkages.
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