蜈蚣碱性蛋白SSmp-d的分离纯化及其部分理化性质的鉴定  

Isolation,Purification and Characterization of SSmp d from Scolopendra subspinipes mutilans L.Koch

作　　者：许鸣镝[1,2] 柳雪枚[1,2] 

机构地区：[1]中国医学科学院 [2]中国协和医科大学药物研究所

出　　处：《生物化学杂志》1997年第5期586-591,共6页

基　　金：国家自然科学基金

摘　　要：少棘巨蜈蚣（ＳｃｏｌｏｐｅｎｄｒａｓｕｂｓｐｉｎｉｐｅｓｍｕｔｉｌａｎｓＬ．Ｋｏｃｈ）经９５％乙醇脱脂后，再经４℃水冷渗，水提液低温旋转浓缩，冻干，得到的冻干粉先后经过ＳｅｐｈａｄｅｘＧ－２５柱，等电聚焦制备电泳，再经ＳｅｐｈａｄｅｘＧ－１５０柱，ＳｅｐｈａｄｅｘＧ－１００柱，最后经ＨＰＬＣ制备得到一个纯的碱性蛋白，命名为ＳＳｍｐ－ｄ．该蛋白经ＨＰＬＣ、超薄等电聚焦电泳检验是均一的．采用ＨＰＬＣ和Ｐｒｏｔｅｉｎ－ＰａｋＴＭ１２５柱测定其分子量为２４．６４ｋＤ．ＩＥＦ－ＨＰＣＥ显示其等电点为９．２７．氨基酸分析表明ＳＳｍｐ－ｄ含较多的Ａｒｇ、Ｌｙｓ等碱性氨基酸，另外还含有较多的Ａｌａ、Ｌｅｕ．使用蛋白质自动序列分析仪测定了ＳＳｍｐ－ｄＮ端的１１个氨基酸，序列为ＮＨ３＋－Ａｓｐ－Ｖａｌ－Ａｓｎ－Ｐｈｅ－Ａｒｇ－Ｌｅｕ－Ｓｅｒ－Ｇｌｙ－Ａｌａ－Ａｓｐ－Ｐｒｏ．A new protein,nominated as SSmp d,has been purified from the water extract of Scolopendra subspinipes mutilans L.Koch. The fraction cotaining SSmp d was separatied from the extract by gel filtration on a Sephadex G 25 column,followed by preparative isoelectric focusing.The new protein purified from the fraction by gel filtration on a Sephadex G 150 column,subsequently on a Sephadex G 100 column,and finally by HPLC on a Protein Pak TM 125 column showed a state of homogeneity on HPLC and IEF PAGE.The isoelectric point of SSmp d determined by IEF HPCE is pH 9 27.Its molecular weight was estimated to be about 24 6 kD by HPLC. Amino acid analysis of SSmp d showed that it was rich in Ala and Leu.Eleven amino acids of the N terminal sequence determined by Edman degradation were proved to be NH + 3 Asp Val Asn Phe Arg Leu Ser Gly Ala Asp Pro.

关 键 词：中药化学 蜈蚣 碱性蛋白 分离 纯化 理化性质 

分 类 号：R284.2[医药卫生—中药学] R282.740.3[医药卫生—中医学]