厚壳贻贝Mytilus coruscus足丝盘及足丝的多巴分析(英文)  

Detection of Dihydroxyphenylalanine in Native Foot Proteins from Mytilus coruscus Byssus

作　　者：李楠楠[1] 石戈[1] 杨宵旭[2] 王智平[1] 廖智[1] 

机构地区：[1]浙江海洋学院海洋科学学院海洋生物资源及分子工程实验室,浙江舟山316004 [2]湖南师范大学生命科学学院蛋白质化学及发育生物学教育部重点实验室,长沙410081

出　　处：《中国生物化学与分子生物学报》2011年第2期148-153,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：Supported by Research and Agricultural Project of Science and Technology Department of Zhejiang Province(No.2008C22026);Key Project of Education Department of Zhejiang Province(No.20070430)~~

摘　　要：多巴(3,4-l-dihydroxyphenylalanine,DOPA)是贻贝足丝粘附蛋白中的一种特殊的氨基酸,由酪氨酸经羟化后生成,与贻贝足丝粘附蛋白的强粘附性能具有直接联系.目前,已鉴定的多种贻贝足丝蛋白序列中均发现有不同含量的DOPA存在.蛋白中DOPA的定量检测对于了解DOPA在蛋白粘附中的作用以及粘附蛋白的粘附机理具有重要意义.为了解厚壳贻贝足丝盘和足丝纤维中DOPA的含量以及DOPA与厚壳贻贝足丝蛋白的粘附性能之间的关系,利用氮蓝四唑(nitrobluetetrazolium,NBT)对DOPA的特异染色原理,结合蛋白质电泳、组织化学染色以及玻璃包被实验,分析和比较了厚壳贻贝足丝不同部位的DOPA含量、蛋白组成特点以及粘附能力.NBT可以在不水解蛋白质的情况下进行DOPA的定量分析,且与DOPA的氧化状态无关.研究结果表明,厚壳贻贝足丝盘中蛋白的DOPA含量明显高于足丝,两者DOPA含量相差2.33倍;蛋白质电泳结果表明,厚壳贻贝足丝盘和足丝纤维的蛋白质组成差异较大,足丝盘中的蛋白质分子量范围明显低于足丝纤维;玻璃包被实验结合POQuest软件半定量分析表明,厚壳贻贝足丝盘蛋白的粘附能力明显高于足丝纤维,且蛋白在玻璃表面的吸附能力与DOPA含量具有相关性.通过以上研究,建立了一种蛋白中DOPA的定量分析方法并初步了解了厚壳贻贝足丝盘和足丝纤维的蛋白组成、DOPA含量以及粘附性能之间的关系,为深入了解贻贝足丝的粘附性能、粘附机制以及开发新型生物粘合剂奠定了基础.The amino acid 3,4-l-dihydroxyphenylalanine （DOPA） can be detected in a variety of foot proteins in mussel. The quantification of DOPA in proteins is essential to understand its role in mussel adhesion. DOPA can be detected with considerable sensitivity with nitroblue tetrazolium /Glycinate（NBT/Glycinate） staining. The Mytilus coruscus byssal proteins were experimented for the detections of the DOPA content,protein distribution,and adhesive ability of foot proteins from thread and plaque with electrophoresis and coating assay as well. We found that the average DOPA content in proteins of plaques was about 2. 33 times higher than that of thread. The adhesive proteins of byssal plaque contained more DOPA and low molecular weight contents than those from the thread with the intensity difference about four folds as determined by PQuest semi-quantitative analysis. The coating ability on glass of the plaque proteins was greater than that of the thread proteins,which correlated with the difference in the protein composition. Our data may help to understand the functional diversity of adhesive proteins in mussel adhesion.

关 键 词：MYTILUS coruscus 多巴(DOPA) 足丝 足丝纤维 足丝盘 

分 类 号：Q78[生物学—分子生物学]