The Perpendicular Orientation of Amphotericin B Methyl Ester in Hydrated Lipid Bilayers Supports the Barrel-Stave Model

Tomoya Yamamoto, Yuichi Umegawa, Masaki Yamagami, Taiga Suzuki, Hiroshi Tsuchikawa, Shinya Hanashima, Nobuaki Matsumori, Michio Murata

Research output: Contribution to journalArticle

Abstract

The clinically important antibiotic amphotericin B (AmB) is a membrane-active natural product that targets membrane sterol. The antimicrobial activity of AmB is generally attributed to its membrane permeabilization, which occurs when a pore is formed across a lipid bilayer. In this study, the molecular orientation of AmB was investigated using solid-state nuclear magnetic resonance (NMR) to better understand the mechanism of antifungal activity. The methyl ester of AmB (AME) labeled with NMR isotopes, d3-AME, and its fluorinated and/or 13C-labeled derivatives were prepared. All of the AmB derivatives showed similar membrane-disrupting activities and ultraviolet spectra in phospholipid liposomes, suggesting that their molecular assemblies in membranes closely mimic those of AmB. Solid-state 2H NMR measurements of d3-AME in a hydrated membrane showed that the mobility of AME molecules depends on concentration and temperature. At a 1:5:45 AME:Erg:dimyristoylphosphatidylcholine ratio, AME became sufficiently mobilized to observe the motional averaging of quadrupole coupling. On the basis of the rotational averaging effect of 19F chemical shift anisotropy, 2H quadrupolar splitting, and 13C-19F dipolar coupling of 14β-F-AMEs, we deduced that the molecular axis of AME is predominantly parallel to the normal of a lipid bilayer. This result supports the barrel-stave model as a molecular assembly of AmB in membranes. ©

Original languageEnglish
Pages (from-to)2282-2291
Number of pages10
JournalBiochemistry
Volume58
Issue number17
DOIs
Publication statusPublished - Apr 30 2019

Fingerprint

Lipid bilayers
Lipid Bilayers
Amphotericin B
Membranes
Magnetic Resonance Spectroscopy
Nuclear magnetic resonance
Magnetic resonance measurement
Dimyristoylphosphatidylcholine
Derivatives
methylamphotericin B
Molecular orientation
Anisotropy
Chemical shift
Sterols
Biological Products
Liposomes
Isotopes
Phospholipids
Anti-Bacterial Agents
Molecules

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Yamamoto, T., Umegawa, Y., Yamagami, M., Suzuki, T., Tsuchikawa, H., Hanashima, S., ... Murata, M. (2019). The Perpendicular Orientation of Amphotericin B Methyl Ester in Hydrated Lipid Bilayers Supports the Barrel-Stave Model. Biochemistry, 58(17), 2282-2291. https://doi.org/10.1021/acs.biochem.9b00180

The Perpendicular Orientation of Amphotericin B Methyl Ester in Hydrated Lipid Bilayers Supports the Barrel-Stave Model. / Yamamoto, Tomoya; Umegawa, Yuichi; Yamagami, Masaki; Suzuki, Taiga; Tsuchikawa, Hiroshi; Hanashima, Shinya; Matsumori, Nobuaki; Murata, Michio.

In: Biochemistry, Vol. 58, No. 17, 30.04.2019, p. 2282-2291.

Research output: Contribution to journalArticle

Yamamoto, T, Umegawa, Y, Yamagami, M, Suzuki, T, Tsuchikawa, H, Hanashima, S, Matsumori, N & Murata, M 2019, 'The Perpendicular Orientation of Amphotericin B Methyl Ester in Hydrated Lipid Bilayers Supports the Barrel-Stave Model', Biochemistry, vol. 58, no. 17, pp. 2282-2291. https://doi.org/10.1021/acs.biochem.9b00180
Yamamoto, Tomoya ; Umegawa, Yuichi ; Yamagami, Masaki ; Suzuki, Taiga ; Tsuchikawa, Hiroshi ; Hanashima, Shinya ; Matsumori, Nobuaki ; Murata, Michio. / The Perpendicular Orientation of Amphotericin B Methyl Ester in Hydrated Lipid Bilayers Supports the Barrel-Stave Model. In: Biochemistry. 2019 ; Vol. 58, No. 17. pp. 2282-2291.
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