Identification of a novel ubiquitin binding site of STAM1 VHS domain by NMR spectroscopy

• Published in: FEBS letters 2009-01, Vol.583 (2), p.287-292
• Main Authors: Hong, Yoon-Hun, Ahn, Hee-Chul, Lim, Jongsoo, Kim, Hong-Man, Ji, Hye-Young, Lee, Seunga, Kim, Ji-Hun, Park, Eun Young, Song, Hyun Kyu, Lee, Bong-Jin
• Format: Article
• Language: English
• Subjects: 1,4-dithiothreitol; 2,2-dimethyl-2-silapentane-5-sulfonic acid; Adaptor Proteins, Signal Transducing - chemistry; Adaptor Proteins, Signal Transducing - genetics; Adaptor Proteins, Signal Transducing - metabolism; Amino Acid Sequence; Binding Sites; chemical shift index; Chemical shift perturbation; coupling of ubiquitin conjugation to endoplasmic reticulum degradation; CSI; CUE; DSS; DTT; endosomal sorting complex required for transport; Endosomal Sorting Complexes Required for Transport; equilibrium dissociation constant; ESCRT; GAT; GGA; GGAs and TOM; glutathione S-transferase; golgi-localizing/γ-adaptin ear homology domain/ADP-ribosylation factor-binding protein; GST; hepatocyte growth factor-regulated substrate; heteronuclear single quantum correlation spectroscopy; Hrs; HSQC; Humans; IPTG; isopropyl β-d-1-thiogalactopyranoside; K d; MIU; Molecular Sequence Data; motif interacting with ubiquitin; multivesicular body; MVB; NMR; NMR spectroscopy; NOE; nuclear magnetic resonance; Nuclear Magnetic Resonance, Biomolecular; nuclear overhauser enhancement; PDB; phenylmethanesulphonylfluoride; Phosphoproteins - chemistry; Phosphoproteins - genetics; Phosphoproteins - metabolism; PMSF; protein data bank; Protein Interaction Domains and Motifs - genetics; Protein Interaction Mapping; Protein Structure, Secondary; Protein–protein interaction; SH3; signal-transducing adapter molecule; Src homology domain 3; Src-activating signaling molecule; Srcasm; STAM; STAM1 VHS domain; target of Myb; TGN; Tom; transgolgi network; Tryptophan - metabolism; UBA; Ubiquitin - chemistry; Ubiquitin - genetics; Ubiquitin - metabolism; ubiquitin associated domain; ubiquitin interacting motif; Ubiquitin recognition; UIM; VHS; Vps27/Hrs/Stam
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/j.febslet.2008.12.034

Interaction between the signal-transducing adapter molecule 1 (STAM1) Vps27/Hrs/Stam (VHS) domain and ubiquitin was investigated by nuclear magnetic resonance (NMR) spectroscopy. NMR evidence showed that the structure of STAM1 VHS domain resembles that of other VHS domains, especially the homologous domain of STAM2. We found that the VHS domain binds to ubiquitin via its hydrophobic patch consisting of N-terminus of helix 2 and C-terminus of helix 4 in which Trp26 on helix 2 plays a pivotal role in the binding. The binding between VHS and ubiquitin seems to be very similar to that between ubiquitin associated domain (UBA) and ubiquitin, however, the direction of α-helices involved in the ubiquitin binding is opposite. Here, we propose a novel ubiquitin binding site and the manner of ubiquitin recognition of the STAM1 VHS domain. MINT- 6804185: STAM1 (uniprotkb: Q92783) binds (MI: 0407) to ubiquitin (uniprotkb: P62988) by nuclear magnetic resonance (MI: 0077)