Localization of clavulones, prostanoids with antitumor activity, within the Okinawan soft coral Clavularia viridis (Alcyonacea, Clavulariidae): preparation of a high-purity Symbiodinium fraction using a protease and a detergent.
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 E, 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.
Tokyo University of Pharmacy and Life Science, Hachioji, Tokyo 192-0392, Japan.
Naoko
N
Hashimoto
Shoko
S
Fujiwara
Kinzo
K
Watanabe
Kazuo
K
Iguchi
Mikio
M
Tsuzuki
D000818:Animals; D037421:Anthozoa / Q000737:chemistry*; D000970:Antineoplastic Agents / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D002855:Chromatography, Thin Layer; D015394:Molecular Structure; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000737:chemistry*; D011453:Prostaglandins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D011454:Prostaglandins A / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000378:metabolism*; D012995:Solubility
D000970:Antineoplastic Agents; D011453:Prostaglandins; D011454:Prostaglandins A; C046249:clavulones; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40; D012697:Serine Endopeptidases; C028734:protease E