脉红螺血蓝蛋白的碳水化合物部分

Carbohydrate moieties of molluscan Rapana venosa hemocyanin.

作者信息

Dolashka-Angelova Pavlina, Beck A, Dolashki A, Stevanovic S, Beltramini M, Salvato B, Hristova R, Velkova L, Voelter W

机构信息

Institute of Organic Chemistry, Bulgarian Academy of Sciences, Sofia, Bulgaria.

出版信息

Micron. 2004;35(1-2):101-4. doi: 10.1016/j.micron.2003.10.032.

DOI:10.1016/j.micron.2003.10.032

PMID:15036306

Abstract

Using Zn2+ ions as new method, several FUs have been isolated from molluscan Hc Rapana venosa without formation of non-functional proteolytic side products. N-terminal sequences of these fragments in comparison with FUS from other gastropodan Hcs show a very high degree of structural identity. Four Fus, purified from enzyme-treated structural subunits RvH1 and RvH2 (RvH1-a, RvH1-f, RvH2-a and RvH2-e) show identical N-terminal sequences compared to fragments isolated after treatment with Zn2+ ions. However, in some cases trypsin cleaves RvH chains at different positions if compared to the Zn2+ treatment. To analyze the oligosaccharide composition of two FUS from the first structural subunit of Rapana Hc, RvH1-a and RvH1-f, several techniques were applied: capillary electrophoresis, MALDI-MS, ESI-MS in combination with glycosidase digestions. On basis of these results and the determined amino acid sequence two N-linkage sites were identified in the FU RvH1-a, but only one in the FU RvH1-f.

摘要

采用锌离子作为新方法，已从软体动物脉红螺的血蓝蛋白中分离出几种功能单元，且未形成无功能的蛋白水解副产物。与其他腹足纲动物血蓝蛋白的功能单元相比，这些片段的N端序列显示出非常高的结构同一性。从经酶处理的结构亚基RvH1和RvH2（RvH1-a、RvH1-f、RvH2-a和RvH2-e）中纯化得到的四种功能单元，与用锌离子处理后分离得到的片段相比，显示出相同的N端序列。然而，在某些情况下，与锌离子处理相比，胰蛋白酶在不同位置切割RvH链。为了分析脉红螺血蓝蛋白第一个结构亚基中的两种功能单元RvH1-a和RvH1-f的寡糖组成，应用了几种技术：毛细管电泳、基质辅助激光解吸电离质谱、电喷雾电离质谱并结合糖苷酶消化。基于这些结果和测定的氨基酸序列，在功能单元RvH1-a中鉴定出两个N-连接位点，但在功能单元RvH1-f中仅鉴定出一个。