Shiomi Kazuo, Midorikawa Sayaka, Ishida Masami, Nagashima Yuji, Nagai Hiroshi
Department of Food Science and Technology, Tokyo University of Marine Science and Technology, Konan-4, Minato-ku, 108-8477, Japan.
Toxicon. 2004 Oct;44(5):499-506. doi: 10.1016/j.toxicon.2004.06.012.
Two lethal factors (named plancitoxins I and II for major and minor toxins, respectively) with the same LD50 (i.v. injection into mice) of 140 microg/kg were purified from spines of the crown-of-thorns starfish Acanthaster planci. Injection of a sublethal dose of plancitoxin I or II into mice remarkably elevated serum levels of glutamic oxaloacetic transaminase and glutamic pyruvic transaminase, demonstrating that both toxins are potently hepatotoxic. Analysis by SDS-PAGE revealed that both plancitoxins are composed of two subunits (alpha-subunit of 10 kDa and beta-subunit of 27 kDa) bridged by a disulfide bond. Based on the determined N-terminal amino acid sequences of alpha- and beta-subunits, the full-length cDNA (1820 bp) encoding plancitoxin I was cloned by RT-PCR, 3'-RACE and 5'-RACE. alpha-Subunit (92 amino acid residues) and beta-subunit (240 residues) are coded in this order by the same cDNA. Interestingly, the deduced amino acid sequence of plancitoxin I showed 40-42% homologies with mammalian deoxyribonucleases II (DNases II). In addition, plancitoxin I exhibited DNA degrading activity with an optimum pH of 7.2. Plancitoxin I is the first example of toxic DNases II whose structures have been elucidated.
从棘冠海星的刺中纯化出了两种致死因子(分别将主要毒素和次要毒素命名为刺参毒素I和II),它们静脉注射到小鼠体内的半数致死剂量(LD50)相同,均为140微克/千克。向小鼠注射亚致死剂量的刺参毒素I或II后,血清中的谷氨酸草酰乙酸转氨酶和谷氨酸丙酮酸转氨酶水平显著升高,表明这两种毒素都具有很强的肝毒性。SDS-PAGE分析显示,两种刺参毒素均由两个亚基(10 kDa的α亚基和27 kDa的β亚基)通过二硫键连接而成。根据所确定的α亚基和β亚基的N端氨基酸序列,通过RT-PCR、3'-RACE和5'-RACE克隆了编码刺参毒素I的全长cDNA(1820 bp)。α亚基(92个氨基酸残基)和β亚基(240个残基)由同一cDNA按此顺序编码。有趣的是,刺参毒素I的推导氨基酸序列与哺乳动物脱氧核糖核酸酶II(DNases II)具有40-42%的同源性。此外,刺参毒素I表现出DNA降解活性,最适pH为7.2。刺参毒素I是首个结构已被阐明的有毒DNases II实例。