5KHY

Crystal structure of oxime-linked K6 diubiquitin

Summary for 5KHY

• Entry DOI: 10.2210/pdb5khy/pdb
• Descriptor: Polyubiquitin-B, ZINC ION, ETHANOLAMINE, ... (4 entities in total)
• Functional Keywords: ubiquitin, oxime, signaling protein
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 2
• Total formula weight: 17541.45

Authors

Stanley, M.,Virdee, S. (deposition date: 2016-06-16, release date: 2016-09-14, Last modification date: 2023-11-15)

Primary citation

Stanley, M.,Virdee, S.
Genetically Directed Production of Recombinant, Isosteric and Nonhydrolysable Ubiquitin Conjugates.
Chembiochem, 17:1472-1480, 2016

PubMed Abstract: 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 the general production of nonhydrolysable ubiquitin conjugates of recombinant origin by bioorthogonal oxime ligation. This was exemplified by the preparation of nonhydrolysable versions of diubiquitin, polymeric ubiquitin chains and ubiquitylated SUMO. The conjugates exhibited unrivalled isostery with the native isopeptide bond, as inferred from structural and biophysical characterisation. Furthermore, the conjugates functioned as nanomolar inhibitors of deubiquitylating enzymes and were recognised by linkage-specific antibodies. This technology should provide a versatile platform for the development of powerful tools for studying deubiquitylating enzymes and for elucidating the cellular roles of diverse polyubiquitin linkages.

PubMed: 27197715
DOI: 10.1002/cbic.201600138

Experimental method

X-RAY DIFFRACTION (3.501 Å)