Solution-state NMR structural ensemble of human Tsg101 UEV in complex with tenatoprazole
Tsg101 chaperone function revealed by HIV-1 assembly inhibitors.
GAVSESQLKKMVSKYKYRDLTVRETVNVITLYKDLKPVLDSYVFNDGSSRELMNLTGTIPVPYRGNTYNIPICLWLLDTY PYNPPICFVKPTSSMTIKTGKHVDANGKIYLPYLHEWKHPQSDLLGLIQVMIVVFGDEPPVFSRP
0.6 mM [U-98% 13C; U-98% 15N] Tsg101, 0.6 mM Tenatoprazole, 20 mM potassium phosphate, 50 mM sodium chloride, 93% H2O/7% D2O
Tenatoprazole (dissolved in DMSO) was allowed to react covalently with Tsg101, which was followed by buffer exchange to remove excess DMSO and tenatoprazole.
0.6 mM [U-13C; U-15N] Tsg101, 0.6 mM Tenatoprazole, 20 mM potassium phosphate, 50 mM sodium chloride, 100% D2O
13C-NOESY experiments were carried out using this sample.
Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
Schwieters, Kuszewski, Tjandra and Clore
Liquid-chromatography mass spectrometry was used to confirm that 4N1 is attached to Tsg101 via a disulfide bond. NMR spectroscopy also confirmed this.
ESCRT-I, inhibitor, cell cycle, cell cycle-inhibitor complex
MAVSESQLKKMVSKYKYRDLTVRETVNVITLYKDLKPVLDSYVFNDGSSRELMNLTGTIPVPYRGNTYNIPICLWLLDTY PYNPPICFVKPTSSMTIKTGKHVDANGKIYLPYLHEWKHPQSDLLGLIQVMIVVFGDEPPVFSRP