PDBsum entry 5khy

Signaling protein

PDB id

5khy

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Contents

			Protein chains

					73 a.a.


					75 a.a.

			Ligands

					ETA

			Metals

					_ZN ×10

PDB id:

5khy

Links

Name:

Signaling protein

Title:

Crystal structure of oxime-linked k6 diubiquitin

Structure:

Polyubiquitin-b. Chain: a. Engineered: yes. Mutation: yes. Polyubiquitin-b. Chain: b. Engineered: yes. Mutation: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: ubb. Expressed in: bacteria. Expression_system_taxid: 2. Expression_system_taxid: 2

Resolution:

3.50Å

R-factor:

0.295

R-free:

0.331

Authors:

M.Stanley,S.Virdee

Key ref:

M.Stanley and S.Virdee (2016). Genetically Directed Production of Recombinant, Isosteric and Nonhydrolysable Ubiquitin Conjugates. Chembiochem, 17, 1472-1480. PubMed id: 27197715

Date:

16-Jun-16

Release date:

14-Sep-16

PROCHECK

	Headers

	References

Protein chain

P0CG47 (UBB_HUMAN) - Polyubiquitin-B from Homo sapiens

Seq: Struc:					229 a.a. 73 a.a.*

Protein chain

P0CG47 (UBB_HUMAN) - Polyubiquitin-B from Homo sapiens

Seq: Struc:					229 a.a. 75 a.a.

Key:

Secondary structure

* PDB and UniProt seqs differ at 1 residue position (black cross)

	Chembiochem 17:1472-1480 (2016)
PubMed id: 27197715

Genetically Directed Production of Recombinant, Isosteric and Nonhydrolysable Ubiquitin Conjugates.
M.Stanley, S.Virdee.

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.