经冻融法提取,丙酮沉淀,SP Sephadex C 25强离子交换和HPLC ODS C18反相色谱层析,从海葵Actinia cari中提纯到两个新的毒素多肽,命名为RSAPⅠ和RSAPⅡ,经SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)和基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定RSAPⅠ和RSAPⅡ均为单一成分,相对分子质量分别为5 008和4 992.初步钠通道药理学实验表明,RSAPⅠ和RSAPⅡ对豚鼠心室肌细胞钠通道表现出活性,RSAPⅠ使钠离子内流增加,起开放作用;RSAPⅡ使钠离子内流减少,起阻滞作用.二者的圆二色谱均表现为100% β转角.
Two toxin polypeptides (designated RSAPⅠ and RSAPⅡ) have been isolated from Sea anemone Actinia cari. Toxins isolation was accomplished by freezing-thawing extraction, acetone fractional precipitation, cation exchange chromagraphy on SP Sephadex C 25 and reverse phase HPLC on ODS C18.They were found to be homogenous by SDS-PAGE and MALDI-TOF-MS, and their relative molecular masses are 5 008 and 4 992, respectively. They are found to be active on sodium channel in Guinea pig cardiac cell, RSAPI, increasing inward Na(+) currents, opens the sodium channel; RSAPⅡ, decreasing inward Na(+) currents, inhibits the opening of the sodium channel. The circular dichroic spectra of these two polypeptides are 100% β turn.
禚如朋,付宏征,仲崇波,王晓良,林文翰,张礼和.海葵毒素多肽的分离和初步表征[J].生物化学与生物物理进展,2001,28(4):514-518
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