Interaction between the marine sponge cyclic peptide theonellamide a and sterols in lipid bilayers as viewed by surface plasmon resonance and solid-state 2H nuclear magnetic resonance

Rafael Atillo Espiritu, Nobuaki Matsumori, Michio Murata, Shinichi Nishimura, Hideaki Kakeya, Shigeki Matsunaga, Minoru Yoshida

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Theonellamides (TNMs) are members of a distinctive family of antifungal and cytotoxic bicyclic dodecapeptides isolated from the marine sponge Theonella sp. Recently, it has been shown that TNMs recognize 3β-hydroxysterol- containing membranes, induce glucan overproduction, and damage cellular membranes. However, to date, the detailed mode of sterol binding at a molecular level has not been determined. In this study, to gain insight into the mechanism of sterol recognition of TNM in lipid bilayers, surface plasmon resonance (SPR) experiments and solid-state deuterium nuclear magnetic resonance (2H NMR) measurements were performed on theonellamide A (TNM-A). SPR results revealed that the incorporation of 10 mol % cholesterol or ergosterol into 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) membranes significantly enhances the affinity of the peptide for the membrane, particularly in the initial binding to the membrane surface. These findings, together with the fact that binding of TNM-A to epicholesterol (3α-cholesterol)-containing liposomes and pure POPC liposomes was comparably weak, confirmed the preference of the peptide for the 3β-hydroxysterol-containing membranes. To further establish the formation of the complex of TNM-A with 3β-hydroxysterols in lipid bilayers, solid-state 2H NMR measurements were conducted using deuterium-labeled cholesterol, ergosterol, or epicholesterol. The 2H NMR spectra showed that TNM-A significantly inhibits the fast rotational motion of cholesterol and ergosterol, but not epicholesterol, therefore verifying the direct complexation between TNM-A and 3β-hydroxysterols in lipid bilayers. This study demonstrates that TNM-A directly recognizes the 3β-OH moiety of sterols, which greatly facilitates its binding to bilayer membranes.

Original languageEnglish
Pages (from-to)2410-2418
Number of pages9
JournalBiochemistry
Volume52
Issue number14
DOIs
Publication statusPublished - Apr 9 2013
Externally publishedYes

Fingerprint

Cyclic Peptides
Lipid bilayers
Surface Plasmon Resonance
Lipid Bilayers
Surface plasmon resonance
Porifera
Sterols
Magnetic Resonance Spectroscopy
Cholesterol
Nuclear magnetic resonance
Membranes
Ergosterol
Deuterium
Liposomes
Theonella
Peptides
Glucans
Complexation
theonellamide A

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Interaction between the marine sponge cyclic peptide theonellamide a and sterols in lipid bilayers as viewed by surface plasmon resonance and solid-state 2H nuclear magnetic resonance. / Espiritu, Rafael Atillo; Matsumori, Nobuaki; Murata, Michio; Nishimura, Shinichi; Kakeya, Hideaki; Matsunaga, Shigeki; Yoshida, Minoru.

In: Biochemistry, Vol. 52, No. 14, 09.04.2013, p. 2410-2418.

Research output: Contribution to journalArticle

Espiritu, Rafael Atillo ; Matsumori, Nobuaki ; Murata, Michio ; Nishimura, Shinichi ; Kakeya, Hideaki ; Matsunaga, Shigeki ; Yoshida, Minoru. / Interaction between the marine sponge cyclic peptide theonellamide a and sterols in lipid bilayers as viewed by surface plasmon resonance and solid-state 2H nuclear magnetic resonance. In: Biochemistry. 2013 ; Vol. 52, No. 14. pp. 2410-2418.
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abstract = "Theonellamides (TNMs) are members of a distinctive family of antifungal and cytotoxic bicyclic dodecapeptides isolated from the marine sponge Theonella sp. Recently, it has been shown that TNMs recognize 3β-hydroxysterol- containing membranes, induce glucan overproduction, and damage cellular membranes. However, to date, the detailed mode of sterol binding at a molecular level has not been determined. In this study, to gain insight into the mechanism of sterol recognition of TNM in lipid bilayers, surface plasmon resonance (SPR) experiments and solid-state deuterium nuclear magnetic resonance (2H NMR) measurements were performed on theonellamide A (TNM-A). SPR results revealed that the incorporation of 10 mol {\%} cholesterol or ergosterol into 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) membranes significantly enhances the affinity of the peptide for the membrane, particularly in the initial binding to the membrane surface. These findings, together with the fact that binding of TNM-A to epicholesterol (3α-cholesterol)-containing liposomes and pure POPC liposomes was comparably weak, confirmed the preference of the peptide for the 3β-hydroxysterol-containing membranes. To further establish the formation of the complex of TNM-A with 3β-hydroxysterols in lipid bilayers, solid-state 2H NMR measurements were conducted using deuterium-labeled cholesterol, ergosterol, or epicholesterol. The 2H NMR spectra showed that TNM-A significantly inhibits the fast rotational motion of cholesterol and ergosterol, but not epicholesterol, therefore verifying the direct complexation between TNM-A and 3β-hydroxysterols in lipid bilayers. This study demonstrates that TNM-A directly recognizes the 3β-OH moiety of sterols, which greatly facilitates its binding to bilayer membranes.",
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T1 - Interaction between the marine sponge cyclic peptide theonellamide a and sterols in lipid bilayers as viewed by surface plasmon resonance and solid-state 2H nuclear magnetic resonance

AU - Espiritu, Rafael Atillo

AU - Matsumori, Nobuaki

AU - Murata, Michio

AU - Nishimura, Shinichi

AU - Kakeya, Hideaki

AU - Matsunaga, Shigeki

AU - Yoshida, Minoru

PY - 2013/4/9

Y1 - 2013/4/9

N2 - Theonellamides (TNMs) are members of a distinctive family of antifungal and cytotoxic bicyclic dodecapeptides isolated from the marine sponge Theonella sp. Recently, it has been shown that TNMs recognize 3β-hydroxysterol- containing membranes, induce glucan overproduction, and damage cellular membranes. However, to date, the detailed mode of sterol binding at a molecular level has not been determined. In this study, to gain insight into the mechanism of sterol recognition of TNM in lipid bilayers, surface plasmon resonance (SPR) experiments and solid-state deuterium nuclear magnetic resonance (2H NMR) measurements were performed on theonellamide A (TNM-A). SPR results revealed that the incorporation of 10 mol % cholesterol or ergosterol into 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) membranes significantly enhances the affinity of the peptide for the membrane, particularly in the initial binding to the membrane surface. These findings, together with the fact that binding of TNM-A to epicholesterol (3α-cholesterol)-containing liposomes and pure POPC liposomes was comparably weak, confirmed the preference of the peptide for the 3β-hydroxysterol-containing membranes. To further establish the formation of the complex of TNM-A with 3β-hydroxysterols in lipid bilayers, solid-state 2H NMR measurements were conducted using deuterium-labeled cholesterol, ergosterol, or epicholesterol. The 2H NMR spectra showed that TNM-A significantly inhibits the fast rotational motion of cholesterol and ergosterol, but not epicholesterol, therefore verifying the direct complexation between TNM-A and 3β-hydroxysterols in lipid bilayers. This study demonstrates that TNM-A directly recognizes the 3β-OH moiety of sterols, which greatly facilitates its binding to bilayer membranes.

AB - Theonellamides (TNMs) are members of a distinctive family of antifungal and cytotoxic bicyclic dodecapeptides isolated from the marine sponge Theonella sp. Recently, it has been shown that TNMs recognize 3β-hydroxysterol- containing membranes, induce glucan overproduction, and damage cellular membranes. However, to date, the detailed mode of sterol binding at a molecular level has not been determined. In this study, to gain insight into the mechanism of sterol recognition of TNM in lipid bilayers, surface plasmon resonance (SPR) experiments and solid-state deuterium nuclear magnetic resonance (2H NMR) measurements were performed on theonellamide A (TNM-A). SPR results revealed that the incorporation of 10 mol % cholesterol or ergosterol into 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) membranes significantly enhances the affinity of the peptide for the membrane, particularly in the initial binding to the membrane surface. These findings, together with the fact that binding of TNM-A to epicholesterol (3α-cholesterol)-containing liposomes and pure POPC liposomes was comparably weak, confirmed the preference of the peptide for the 3β-hydroxysterol-containing membranes. To further establish the formation of the complex of TNM-A with 3β-hydroxysterols in lipid bilayers, solid-state 2H NMR measurements were conducted using deuterium-labeled cholesterol, ergosterol, or epicholesterol. The 2H NMR spectra showed that TNM-A significantly inhibits the fast rotational motion of cholesterol and ergosterol, but not epicholesterol, therefore verifying the direct complexation between TNM-A and 3β-hydroxysterols in lipid bilayers. This study demonstrates that TNM-A directly recognizes the 3β-OH moiety of sterols, which greatly facilitates its binding to bilayer membranes.

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