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.
Hashimoto
Naoko
N
Tokyo University of Pharmacy and Life Science, Hachioji, Tokyo 192-0392, Japan.
Fujiwara
Shoko
S
Watanabe
Kinzo
K
Iguchi
Kazuo
K
Tsuzuki
Mikio
M
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