盐离子对次壶腹腺丝蛋白重组模块的影响  

Effect of salt on minor ampullate silk spidroin modules

作　　者：王佳[1] 陈格飞[1] 孟清[1] 

机构地区：[1]东华大学生物科学与技术研究所,上海201620

出　　处：《生物工程学报》2014年第8期1308-1317,共10页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(No.31070698);国家高技术研究发展计划(863计划)(No.2006AA03Z451);上海市科委基础研究重点项目(No.10JC1400300);教育部博士点基金项目(No.20120075110007)资助~~

摘　　要：为研究不同生理环境对蛛丝蛋白组装及成丝的影响,首次以MiSp序列为对象,研究其NTR1SR2CT重组模块在不同种类(浓度)盐离子条件下的聚集和成纤维特性及其在成纤维过程中二级结构的变化。基于大腹园蛛MiSp全长序列构建NTR1SR2CT模块,并在大肠杆菌Escherichia coli BL21中成功表达。借助8 mol/L尿素裂解包涵体进行变性纯化得到NTR1SR2CT重组蛋白。NTR1SR2CT重组蛋白二级结构主要为无规则卷曲(Random coil)或α螺旋(Helix),在自然成丝及冻干过程中部分random coil或helix转变为β折叠(β-sheet),甲醇能促进该转变过程。另外,钾离子和磷酸根离子有利于NTR1SR2CT重组蛋白聚集从而促进丝纤维的形成。研究结果为成丝机理研究奠定了基础,同时也为工业化生产高品质的蛛丝纤维提供了条件。To study the effect of physiological conditions on spidroins, we analyzed NTR 1SR2CT module secondary structure, aggregation and silk-formation influenced by different salts （in different concentration intervals）. According to the full-length Araneus ventricosus MiSp sequence, NTR1SR2CT module was constructed and expressed in Escherichia coli BL21 （DE3）, and the recombinant proteins were purified by denaturation method mediated by 8 mol/L urea. Random coil and helix are the main secondary structures of NTR1SR2CT and could be induced into beta-sheet by drying natively and lyophilization, where methanol can be used as a promoter. Furthermore, potassium and phosphate cations can cause significant NTRISR2CT protein aggregationand silk-formation. The results could be a basis for the determination of silk-formation mechanism, and also useful for industrialized generation of high performance spider silk-like fibers.

关 键 词：次壶腹腺丝蛋白 组装 盐离子 二级结构 丝纤维 

分 类 号：Q51[生物学—生物化学]