title: Modification of bafilomycin structure to efficiently synthesize solid-state NMR probes that selectively bind to vacuolar-type ATPase.
abstract: Bafilomycin (Baf) is one of the most potent inhibitors of vacuolar-type ATPase, which is strongly implicated in age-related diseases. However, the binding site of the antibiotic on the protein remains unclear because of the complexity of the structure of Baf bound to the target subunit in the transmembrane region. For conducting structural studies by applying solid-state NMR, which is one of the most promising methodologies available for structural analysis in membrane system, preparing bioactive fluorinated Baf analogues is essential. In this study two Baf analogues were carefully designed and efficiently synthesized through the convergent coupling of three segments. Biological evaluation revealed that the activity of 24-F-Baf was comparable to that of Baf, indicating its utility as a potential probe for solid-state NMR analysis. By contrast, desmethylated 24-F-Baf exhibited markedly diminished activity. The absence of two methyl groups caused a critical conformational change in the macrocyclic core structure necessary for binding to the target protein.
authors.affiliation: Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043 (Japan), Fax: (+81) 6-6850-5774; ERATO, Lipid Active Structure Project, Japan Science and Technology Agency, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043 (Japan).
authors.forename: Hajime
authors.initials: H
authors.lastname: Shibata
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Tsuchikawa
authors.forename: Tatsuru
authors.initials: T
authors.lastname: Hayashi
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Matsumori
authors.forename: Michio
authors.initials: M
authors.lastname: Murata
authors.forename: Takeo
authors.initials: T
authors.lastname: Usui
mesh_terms: D018942:Macrolides / Q000737:chemistry* / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D011485:Protein Binding; D025262:Vacuolar Proton-Translocating ATPases / Q000378:metabolism*; D014617:Vacuoles / Q000201:enzymology; D015003:Yeasts / Q000201:enzymology
chemical_list: D018942:Macrolides; C040929:bafilomycin A1; D025262:Vacuolar Proton-Translocating ATPases
keywords: V-ATPase; bafilomycin; chemical synthesis; fluorine label; solid-state NMR