Lim, Jongsoo Son, Woo-Sung Park, Joon Kyu Kim, Eunice EunKyeong Lee, Bong-Jin Ahn, Hee-Chul 2024-01-20T17:33:33Z 2024-01-20T17:33:33Z 2021-09-02 2011-02-04 0006-291X https://pubs.kist.re.kr/handle/201004/130647 STAM1 and Hrs are the components of ESCRT-0 complex for lysosomal degradation of membrane proteins is composed of STAM1 Hrs and has multiple ubiquitin binding domains. Here, the solution structure of STAM1 UIM, one of the ubiquitin binding motif, was determined by NMR spectroscopy. The structure of UIM adopts an a-helix with amphipathic nature. The central hydrophobic residues in UIM provides the binding surface for ubiquitin binding and are flanked with positively and negatively charged residues on both sides. The docking model of STAM1 UIM-ubiquitin complex is suggested. In NMR and ITC experiments with the specifically designed mutant proteins, we investigated the ubiquitin interaction of tandem ubiquitin binding domains from STAM1. The ubiquitin binding affinity of the VHS domain and UIM in STAM1 was 52.4 and 94.9 mu M, and 1.5 and 2.2 fold increased, respectively, than the value obtained from the isolated domain or peptide. The binding affinities here would be more physiologically relevant and provide more precise understanding in ESCRT pathway of lysosomal degradation. (c) 2010 Elsevier Inc. All rights reserved. English ACADEMIC PRESS INC ELSEVIER SCIENCE ESCRT MACHINERY RECEPTOR DEGRADATION COOPERATE PROTEINS SYSTEM Solution structure of UIM and interaction of tandem ubiquitin binding domains in STAM1 with ubiquitin Article 10.1016/j.bbrc.2010.12.103 1 BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, v.405, no.1, pp.24 - 30 BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 405 1 24 30 scie scopus 000287336800005 2-s2.0-79551551936 Biochemistry & Molecular Biology Biophysics Biochemistry & Molecular Biology Biophysics Article ESCRT MACHINERY RECEPTOR DEGRADATION COOPERATE PROTEINS SYSTEM Ubiquitin STAM1 ESCRT Lysosomal degradation UIM VHS NMR ITC