烟草丝束蛋白ABD2与微丝的体内体外相互作用  

Tobacco Fimbrin Interacts with Actin Filaments through its Actin-binding Domain 2 in Vitro and in Vivo

作　　者：梁蒙[1] 韩利波[1] 孙海涛[1] 黄璐[1] 陈志玲[1] 

机构地区：[1]首都师范大学生命科学学院,北京100048

出　　处：《中国生物化学与分子生物学报》2014年第9期894-898,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：北京市属高校人才强教计划资助项目(No.PHR201108148);北京市教委科技计划面上项目(No.KM201310028010)~~

摘　　要：微丝骨架在细胞的生命活动中具有重要的功能,而其动态的解聚聚合特性是其实现功能的前提.丝束蛋白(fimbrin/plastin)做为微丝结合蛋白质,是微丝骨架的重要调控因子之一,含有2个肌动蛋白结合结构域,目前对其结合微丝的机制并不清楚.本文以烟草丝束蛋白的肌动蛋白结合结构域2(NtFAbd2)为研究对象,通过原核细胞表达纯化NtFAbd2,利用体外沉淀法分析发现,NtFAbd2能够与微丝结合;利用激光共聚焦扫描显微镜分析发现,在烟草BY-2悬浮细胞内,NtAbd2-GFP与微丝共分布,这些结果为深入分析植物丝束蛋白的作用机制提供了新的数据.The actin cytoskeleton is important for numerous cellular processes and functions dynamically. Fimbrins belong to a family of F-actin cross-linking proteins and control F-actin dynamics. Each member of the fimbrin/plastin family contains two highly conserved actin- binding domains. In present study, recombinant NtFAbd2 proteins were expressed in E. coli and purified using a Ni-NTA agarose affinity column. High-speed co-sedimentation assays demonstrated that NtFAbd2 interact with F-actin in vitro. Laser scanning confocal microscopy revealed that NtFAbd2-GFP fusions labeled filamentous structures and coloealized with rhodamine-phalloidin labeled actin filaments in tobacco BY-2 suspension cells. These results verified the molecular mechanism of plant fimbrins in the regulation of actin dynamics.

关 键 词：丝束蛋白 肌动蛋白结合结构域2 微丝 烟草BY-2悬浮细胞 

分 类 号：Q291[生物学—细胞生物学]