天然麝香抗炎蛋白质的研究Ⅰ.Mu-a-1的分离纯化及其部分性质的鉴定  

STUDIES ON THE ANTIINFLAMMATORY PROTEIN OF MUSK.Ⅰ.ISOLATION, PURIFICATION AND PROPERTIES

作　　者：柳雪枚[1] 李虹奇 肖宣[1] 于德泉[1] 

机构地区：[1]中国医学科学院药物研究所,北京100050

出　　处：《动物学报》1992年第3期302-308,共7页ACTA ZOOLOGICA SINICA

基　　金："人工麝香"研究课题一部分;受国家科委"六.五"攻关项目资助

摘　　要：天然麝香粉(Musk)经乙醚,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Sephadex G-25,Sephadex G-50,和Sephadex G-100等柱层析,得到抗炎有效部分Mu-a,将此部分再通过DEAE-Sephadex A-25和Sephadex G-150等柱层析,得到一抗炎有效蛋白质,命名为Mu-a-1。经聚丙烯酰胺凝胶电泳及超薄等电聚焦电泳,以高碘酸-schiff试剂及Coomassie R250染色和分子筛层析,均显示一条区带和一均匀吸收峰。以蒽酮试剂法测定Mu-a-1含糖量在40％左右。经氨基酸自动分析仪测定Mu-a-1是由16种氨基酸组成,其中Glu,Asp等酸性氨基酸居多,等电点(PI=3-4),为酸性糖蛋白。最小蛋白质碎片含374个氨基酸残基。用凝胶过滤法估测其分子量在20万左右。药理实验表明Mu-a-1对巴豆油引起小鼠耳部炎症具有明显的抗炎作用,平均7.5mg/kg剂量下,对炎症抑制率为72.3％,P<0.001。An effective method for protein purification and some properties of the antiinflammatory protein from Musk are described for the first time.A water soluble glycoprotein of Musk, named Mu-a-1, was purified by gel filtration on Sephadex G-25 (3cm×84cm), Sephadex G-50 (3cm×50cm), Sephadex G-100 (4cm×50cm) and DEAE-Sephadex A-25 (2cm×46cm) columns as a high yield and electrophoretically pure productMu-a-1 showed a single band on polyacrylamide gel mini Isoelectric focusing electrophoresis and disk polyacrylamide gel electrophoresis.Amino acid composition of Mu-a-1 as determined. The Mu-a-1 consisted of about 16 amino acids, among which acidic amino acid residues (Glu, Asp) were dominant, 23%, basic amino acid residues (Arg, His) were less than 6%, indicating that Mu-a-1 was a strongly acidic protein. Correspondingly, its isoelectric point was about pH = 3-4 as determined by polyacrylamide gel mini Isoelectric focusing electrophoresis.The results of amino acid analyses suggested a minimum protein unit of 374 amino acids.It appeared to be a homogeneous glycoprotein as shown by periodic acid Schiff procedure. The total carbohydrate content was about 40%.The molecular weight of Mu-a-1 was 200000 as determined by gel fitration.Mu-a-1 was relatively stable to heat.

关 键 词：天然麝香 抗炎蛋白 

分 类 号：R284[医药卫生—中药学]