6SYF

Human Ubc9 with covalent isopeptide ligand

Summary for 6SYF

• Entry DOI: 10.2210/pdb6syf/pdb
• Descriptor: SUMO-conjugating enzyme UBC9, ACE-LEU-ARG-LEU-ARG-GLY-CYS, ACE-ILE-LYS-GLN-GLU, ... (4 entities in total)
• Functional Keywords: ubc9, isopeptide, disulfide, ubiquitin, sumo, ligase
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 12
• Total formula weight: 76155.79

Authors

Hofmann, R.,Akimoto, G.,Wucherpfennig, T.G.,Zeymer, C.,Bode, J.W. (deposition date: 2019-09-27, release date: 2020-08-12, Last modification date: 2024-10-16)

Primary citation

Hofmann, R.,Akimoto, G.,Wucherpfennig, T.G.,Zeymer, C.,Bode, J.W.
Lysine acylation using conjugating enzymes for site-specific modification and ubiquitination of recombinant proteins.
Nat.Chem., 12:1008-1015, 2020

PubMed Abstract: Enzymes are powerful tools for protein labelling due to their specificity and mild reaction conditions. Many protocols, however, are restricted to modifications at protein termini, rely on non-peptidic metabolites or require large recognition domains. Here we report a chemoenzymatic method, which we call lysine acylation using conjugating enzymes (LACE), to site-specifically modify folded proteins at internal lysine residues. LACE relies on a minimal genetically encoded tag (four residues) recognized by the E2 small ubiquitin-like modifier-conjugating enzyme Ubc9, and peptide or protein thioesters. Together, this approach obviates the need for E1 and E3 enzymes, enabling isopeptide formation with just Ubc9 in a programmable manner. We demonstrate the utility of LACE by the site-specific attachment of biochemical probes, one-pot dual-labelling in combination with sortase, and the conjugation of wild-type ubiquitin and ISG15 to recombinant proteins.

PubMed: 32929246
DOI: 10.1038/s41557-020-0528-y

Experimental method

X-RAY DIFFRACTION (1.9 Å)