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Identification of a novel ubiquitin binding site of STAM1 VHS domain by NMR spectroscopy |
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| Authors: | Yoon-Hun Hong Jongsoo Lim Hong-Man Kim Seunga Lee Eun Young Park Bong-Jin Lee |
| Affiliation: | a Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, San 56-1, Shillim-dong, Kwanak-gu, Seoul 151-742, Republic of Korea b Advanced Analysis Center, Korea Institute of Science and Technology, 39-1 Hawolgok-dong, Seongbuk-gu, Seoul 136-791, Republic of Korea c Biomolecular Science, University of Science and Technology, 113 Gwahangno, Yuseong-gu, Daejeon 305-333, Republic of Korea d School of Life Sciences and Biotechnology, Korea University, Anam-dong, Seongbuk-gu, Seoul 136-701, Republic of Korea |
| Abstract: | 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.Structured summaryMINT-6804185:STAM1 (uniprotkb:Q92783) binds (MI:0407) to ubiquitin (uniprotkb:P62988) by nuclear magnetic resonance (MI:0077) |
| Keywords: | STAM, signal-transducing adapter molecule VHS, Vps27/Hrs/Stam NMR, nuclear magnetic resonance UBA, ubiquitin associated domain ESCRT, endosomal sorting complex required for transport UIM, ubiquitin interacting motif GAT, GGAs and TOM MVB, multivesicular body SH3, Src homology domain 3 GGA, golgi-localizing/γ-adaptin ear homology domain/ADP-ribosylation factor-binding protein Hrs, hepatocyte growth factor-regulated substrate Tom, target of Myb Srcasm, Src-activating signaling molecule TGN, transgolgi network PDB, protein data bank GST, glutathione S-transferase IPTG, isopropyl β- smallcaps" >d-1-thiogalactopyranoside DTT, 1,4-dithiothreitol PMSF, phenylmethanesulphonylfluoride HSQC, heteronuclear single quantum correlation spectroscopy DSS, 2,2-dimethyl-2-silapentane-5-sulfonic acid CSI, chemical shift index NOE, nuclear overhauser enhancement Kd, equilibrium dissociation constant CUE, coupling of ubiquitin conjugation to endoplasmic reticulum degradation MIU, motif interacting with ubiquitin |
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