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Localization of clavulons, prostanoids with antitumor activity, within the Okinawan soft coral Clavularia viridis (alcyonacea, clavulariidae): Preparation of a high-purity Symbiodinium faction using a protease and a detergent

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Lipids

Abstract

To investigate the localization of clavulones (CV), prostanoids with antitumor activity, in the Okinawan soft coral Clavularia viridis, we developed a method for the isolation of Symbiodinium cells from the coral, i.e., treatment of a coral homogenate with a protease, pronase F, and a detergent, Nonidet P-40. The conditions for the treatment were optimized by monitoring the morphology microscopically and the amount of chlorophyll in the Symbiodinium fraction (SymF) optically. To evaluate the purity of SymF and a Symbiodinium-free coral fraction (CorF), we analyzed them for proteins and lipids using cultivated Symbiodinium as a reference. TLC of lipids revealed that SymF contained a greater amount of glycolipids, whereas CorF comprised mostly phospholipids. SDS-PAGE of proteins in SymF and CorF revealed their distinct profiles. Thus, we could obtain each fraction with high purity; we reached the conclusion that CV and arachidonic acid, their possible precursor, are localized exclusively in the insoluble fraction of host coral cells.

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Abbreviations

AA:

arachidonic acid

CBP:

chlorophyll a-c-binding protein

Chl:

chlorophyll

CorF:

Symbiodinium-free coral fraction

CorHo:

coral homogenate treated with pronase E

CSym:

cultured Symbiodinium

CV:

clavulone(s)

NP-40:

Nonidel P-40

PCP:

peridinin-Chl-binding protein

ppt:

precipitate

sup:

supernatant

SymF:

Symbiodinium fraction

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Authors and Affiliations

  1. CREST, Japan

    Shoko Fujiwara & Mikio Tsuzuki

  2. Tokyo University of Pharmacy and Life Science, 1432-1 Horinouchi, 192-0392, Hachioji, Tokyo, Japan

    Naoko Hashimoto, Shoko Fujiwara, Kinzo Watanabe, Kazuo Iguchi & Mikio Tsuzuki

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  1. Naoko Hashimoto
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  2. Shoko Fujiwara
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  3. Kinzo Watanabe
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  4. Kazuo Iguchi
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  5. Mikio Tsuzuki
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Correspondence to Mikio Tsuzuki.

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Hashimoto, N., Fujiwara, S., Watanabe, K. et al. Localization of clavulons, prostanoids with antitumor activity, within the Okinawan soft coral Clavularia viridis (alcyonacea, clavulariidae): Preparation of a high-purity Symbiodinium faction using a protease and a detergent. Lipids 38, 991–997 (2003). https://doi.org/10.1007/s11745-003-1153-2

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