蚯蚓纤溶酶组分的抗原性分析  

Antigenicity Analysis of Fractions of Earthworm Fibrinolitic Enzyme

作　　者：赵晓瑜[1] 侯艳[1] 高珊[1] 李晓霞[1] 

机构地区：[1]河北大学生命科学学院,保定071002

出　　处：《中国生物化学与分子生物学报》2007年第2期147-153,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：河北省生物工程重点学科资助~~

摘　　要：用热变性蚯蚓纤溶酶(earthworm fibrinolitic enzyme,EFE)组分EfP-0-2、EfP-Ⅰ-1、EfP-Ⅱ-1,以及重组蛋白EfP-Ⅲ-2为抗原,免疫家兔获得了抗血清,利用获得的抗血清对部分组分的免疫学同一性进行了验证.EfP-0、EfP-Ⅰ、EfP-Ⅱ的免疫双扩散反应表明,EfP-Ⅰ和EfP-Ⅱ具有部分免疫学同一性.利用DNAMAN软件对获得的EfP-0、EfP-Ⅰ、EfP-Ⅱ、EfP-Ⅲ的蛋白质序列进行了初步分析,包括整个序列相似性比对、氨基酸组成、蛋白酶消化、抗原肽、疏水性和亲水性轮廓等,并对N端氨基酸序列进行了分析.结果表明,EfP-Ⅰ和EfP-Ⅱ具有部分免疫学同一性是由它们蛋白质序列的组成和结构决定的.Earthworm fibrinolitic enzymes （EFE） were separated from earthworm by affinity chromatography using soybean trypsin inhibitor as a ligand and Sepharose-4B as a cartier. The enzymes had 11-13 protein bands on PAGE. EFE were further separated and purified into 10 fractions, which were divided into four groups： EfP-0, EfP-Ⅰ , EfP-Ⅱ and EfP-Ⅲ, according to N-terminal amino acid sequence. Antisera against EfP-0-2, EfP-Ⅰ-1, EfP-Ⅱ-1 and EfP-Ⅲ-2 were produced by immunizing domestic rabbit with heat denaturalized earthworm fibrinolitic enzyme fractions of EfP-0-2, EfP- Ⅰ -1, EfP-Ⅱ-1 and recombinant protein EfP-Ⅲ-2. The antisera were used to verify the identity of immuno-reaction of some fractions. Immunodiffusion analysis of EfP-0, EfP-Ⅰ and EfP- Ⅱ with these antisera showed that EfP-Ⅰ and EfP-Ⅱ had partial immunoidentity. DNAMAN analysis of EfP-0, EfP-Ⅰ , EfP-Ⅱ and EfP-Ⅲ, including alignment of whole protein sequence similarity, amino acids composition, protein digestion, antigenic peptides, hydrophobicity and hydrophilicity, and amino acid sequence of N-terminal analysis, suggested that the partial immuno-identity of EfP-Ⅰ and EfP-Ⅱ was determined by their sequence composition and structure .

关 键 词：蚯蚓(Eisenia foetida) 纤溶酶 抗原性 免疫反应同一性 抗原肽 

分 类 号：Q55[生物学—生物化学]