毛头鬼伞(Coprinus comatus)中一种碱性蛋白的纯化及其活性  

Purification and Activities of an Alkaline Protein from Mushroom Coprinus comatus

作　　者：吴丽萍[1] 吴祖建[1] 林奇英[1] 谢联辉[1] 

机构地区：[1]福建农林大学植物病毒研究所,福州350002

出　　处：《微生物学报》2003年第6期793-798,共6页Acta Microbiologica Sinica

基　　金：教育部骨干教师资助项目 (2 0 0 0 65 )~~

摘　　要：用离子交换层析 (CM sepharoseFF)和凝胶层析 (SuperdexTM75 )方法 ,从新鲜食用菌毛头鬼伞 (Coprinuscomatus)子实体中分离纯化出一碱性蛋白y3,经SDS PAGE初步确定其分子量约为 1 4 4kD。活性检测结果显示 :当其浓度为 1 2 5 μg mL时 ,对烟草花叶病毒 (TMV)在心叶烟枯斑寄主上的侵染抑制率达 83 0 % ;y3对兔血凝集活性滴度为 2 5,对人血凝集活性滴度为 2 6,其浓度分别为 1 5 62 μg mL和 0 781 μg mL ;利用胃癌细胞株MGC 80 3检测y3体外抗肿瘤活性 ,其IC50 为 1 2 μg mL。y3N 端序列为NRDVAACARFIDDFCDTLTP ,为一新的蛋白序列。在SWISS PORT上登录号为P83477。An Alkaline protein,y3,can be purified from the fruiting bodies of mushroom Coprinus comatus by means of CM-sepharose FF ion-exchange column chromatography and Superdex TM 75 High Resolution molecular sieve chromatography. The protein has a molecular weight of about 14.4kD by SDS-PAGA. Some activities of y3 have been detected,and the result is the following: the inhibition rate against Tobacco Mosaic Virus is 83.0% when the concentration of y3 is 12.5μg/mL. y3 is able to agglutinate rabbit and human erythrocytes at the concentration of 1.562μg/mL and 0.781μg/mL. Using an assay system based on mach cancer cell line MGC-803,y3 was studied for its inhibitory ability against celles multiplication,and the IC_ 50 is 12μg/mL.The N-terminal sequence is NRDVAACARFIDDFCDTLTP,which has no homology with other sequences in Genbank.

关 键 词：分离 纯化 烟草花叶病毒 凝集素 抗肿瘤 毛头鬼伞 食用菌 

分 类 号：Q946[生物学—植物学]