Zykova T A, Monastyrnaia M M, Apalikova O V, Shvets T V, Kozlovskaia E P
Pacific Institute of Bioorganic Chemistry, Far-Eastern Division, Russian Academy of Sciences, Vladivostok, Russia.
Bioorg Khim. 1998 Jul;24(7):509-16.
Two low-molecular cytolytic toxins (RmI and RmII) and four trypsin inhibitors were isolated from the aqueous extract of sea anemone Radianthus macrodactylus. The method of isolation involved precipitation with acetone, gel filtration on acrylex P-4, ion-exchange chromatography on CM-32 cellulose, affinity chromatography on trypsin-binding sepharose 4B, ion exchange chromatography on an Ultrapore TSK CM-3SW column, and reversed phase HPLC on a Silasorb C18 column. RmI, RmII, and JnI inhibitor displayed molecular masses 5100, 6100, and 7100 Da, respectively, when subjected to SDS-PAGE. The isoelectric points were 9.2 and 9.3 for RmI and RmII, respectively. The amino acid composition and N-terminal amino acid residue (glycine) were determined for RmI, RmII, and JnI. Both proteins were nontoxic to mice and crabs. Hemolytic activity was determined to be 25 and 20 HU/mg for RmI and RmII, respectively, and their action on erythrocyte membrane was not inhibited by exogenous sphingomyelin. RmI and RmII exhibited antihistamine activity.
从大触手海葵（Radianthus macrodactylus）的水提取物中分离出两种低分子细胞溶解毒素（RmI和RmII）以及四种胰蛋白酶抑制剂。分离方法包括用丙酮沉淀、在丙烯腈P-4上进行凝胶过滤、在CM-32纤维素上进行离子交换色谱、在胰蛋白酶结合琼脂糖4B上进行亲和色谱、在Ultrapore TSK CM-3SW柱上进行离子交换色谱以及在Silasorb C18柱上进行反相高效液相色谱。当进行SDS-PAGE时，RmI、RmII和JnI抑制剂的分子量分别为5100、6100和7100 Da。RmI和RmII的等电点分别为9.2和9.3。测定了RmI、RmII和JnI的氨基酸组成和N端氨基酸残基（甘氨酸）。这两种蛋白质对小鼠和螃蟹均无毒。RmI和RmII的溶血活性分别测定为25和20 HU/mg，并且它们对红细胞膜的作用不受外源性鞘磷脂的抑制。RmI和RmII表现出抗组胺活性。
