1H, 13C, and 15N resonance assignments of the C-terminal lobe of the human HECT E3 ubiquitin ligase ITCH

• Published in: Biomolecular NMR assignments 2019-04, Vol.13 (1), p.15-20
• Main Authors: Beasley, Steven A., Bardhi, Roela, Spratt, Donald E.
• Format: Article
• Language: English
• Subjects: Apoptosis; Biochemistry; Biological and Medical Physics; Biophysics; C-Terminus; Cell activation; Cell differentiation; Inflammation; Intracellular signalling; Lymphocytes T; Magnetic resonance spectroscopy; NMR; Nuclear magnetic resonance; Physics; Physics and Astronomy; Polymer Sciences; Protein structure; Proteins; Resonance; Transcription factors; Ubiquitin; Ubiquitin-protein ligase
• ISSN: 1874-2718; 1874-270X
• DOI: 10.1007/s12104-018-9843-2

ITCH ( aka Atrophin-1-interacting protein 4) is a prominent member of the NEDD4 HECT ( H omologous to E 6AP C-T erminus) E3 ubiquitin ligase family that regulates numerous cellular functions including inflammatory responses through T-cell activation, cell differentiation, and apoptosis. Known intracellular targets of ITCH-dependent ubiquitylation include receptor proteins, signaling molecules, and transcription factors. The HECT C -terminal lobe of ITCH contains the conserved catalytic cysteine required for the covalent attachment of ubiquitin onto a substrate and polyubiquitin chain assembly. We report here the complete experimentally determined 1 H, 13 C, and 15 N backbone and sidechain resonance assignments for the HECT C -terminal lobe of ITCH (residues 784–903) using heteronuclear, multidimensional NMR spectroscopy. These resonance assignments will be used in future NMR-based studies to examine the role of dynamics and conformational flexibility in HECT-dependent ubiquitylation as well as deciphering the structural and biochemical basis for polyubiquitin chain synthesis and specificity by ITCH.