6XZ1

Conjugate of the HECT domain of HUWE1 with ubiquitin

Summary for 6XZ1

• Entry DOI: 10.2210/pdb6xz1/pdb
• Descriptor: HECT, UBA and WWE domain containing 1, isoform CRA_a, Polyubiquitin-B, PHOSPHATE ION, ... (5 entities in total)
• Functional Keywords: ubiquitin ligase, e3 enzyme, ubiquitin-propargylamide, activity-based probe, ligase
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 4
• Total formula weight: 107826.82

Authors

Liu, B.,Seenivasan, A.,Nair, R.,Chen, D.,Lowe, E.D.,Lorenz, S. (deposition date: 2020-01-31, release date: 2021-08-25, Last modification date: 2024-01-24)

Primary citation

Nair, R.M.,Seenivasan, A.,Liu, B.,Chen, D.,Lowe, E.D.,Lorenz, S.
Reconstitution and Structural Analysis of a HECT Ligase-Ubiquitin Complex via an Activity-Based Probe.
Acs Chem.Biol., 16:1615-1621, 2021

PubMed Abstract: Ubiquitin activity-based probes have proven invaluable in elucidating structural mechanisms in the ubiquitin system by stabilizing transient macromolecular complexes of deubiquitinases, ubiquitin-activating enzymes, and the assemblies of ubiquitin-conjugating enzymes with ubiquitin ligases of the RING-Between-RING and RING-Cysteine-Relay families. Here, we demonstrate that an activity-based probe, ubiquitin-propargylamine, allows for the preparative reconstitution and structural analysis of the interactions between ubiquitin and certain HECT ligases. We present a crystal structure of the ubiquitin-linked HECT domain of HUWE1 that defines a catalytically critical conformation of the C-terminal tail of the ligase for the transfer of ubiquitin to an acceptor protein. Moreover, we observe that ubiquitin-propargylamine displays selectivity among HECT domains, thus corroborating the notion that activity-based probes may provide entry points for the development of specific, active site-directed inhibitors and reporters of HECT ligase activities.

PubMed: 34403242
DOI: 10.1021/acschembio.1c00433

Experimental method

X-RAY DIFFRACTION (2.3 Å)