腹足纲Helix pomatia βc-血清蛋白的功能单位e中碳水化合物的定位和糖基化位置的鉴定(英文)  

Location of The Carbohydrate and Identification of The Glycosylation Sites in Functional Unit e of βc-haemocyanin of The Gastropod Helix Pomatia

作　　者：蔡皓[1] Gielens Constant Preaux Gisele 

机构地区：[1]南京中医药大学江苏省海洋药物研究开发中心,江苏南京210029 [2]Laboratorium voor Biochemie,Katholieke Universiteit te Leuven,Celestijnenlaan 200G,B-3001 Leuven(Heverlee),Belgium

出　　处：《药物生物技术》2004年第5期311-315,共5页Pharmaceutical Biotechnology

摘　　要：对腹足纲软体动物罗曼蜗牛Helixpomatia的氧运输蛋白(βC血清蛋白)的功能单位e中的碳水化合物进行了研究。以该βC血清蛋白为载体,通过限制性胰蛋白酶水解,产生片段ac,ef,以及功能单位d,g,和h。以片段ef降解后的产物为研究对象,通过亲合色谱的分离纯化,得到功能单位e的一个馏分e2。该馏分经还原、吡啶乙基化和胰蛋白酶水解后,通过柱色谱的分离纯化,得到一个含有糖肽的成分。对该糖肽中糖组分和肽序列的分析,最终确定了功能单位e的两个糖基化位置Asn111和Asn387。通过与功能单位d和g的糖基化位置的比较得出结论Asn387为一恒定的糖基化位置。The carbohydrate in functional unit e of β C haemocyanin of the gastropod Helix pomatia was studied. The limited trypsinolysis on tenth molecules of β C haemocyanin of Helix pomatia at pH=8.2 yielded the fragments a c , ef , and the functional units d , g , and h (present as dimers h 2 ). The mixture of functional units e and f after limited fragmentation of fragment ef on storage, were further fractionated with an affinity chromatography in the presence of α methylglucoside, yielded functional unit e 2 . Further separation of trypsinolysis of pyridylethylated functional unit e 2 yielded a fraction I (contain glycopeptides). The analysis of carbohydrate content and the N terminal sequence allowed the location of the carbohydrate and the identification of the glycosylation sites in functional unit e : site A at Asn 111 and site B at Asn 387. From a comparison with the glycosylation sites of Hp d and Hp g , it follows that the site B is a conserved one.

关 键 词：βc-血清蛋白 反相高效液相色谱 气液色谱 DABITC-PITC双偶合法 

分 类 号：Q7[生物学—分子生物学]