鼠肌纤维丝切蛋白CFL2互作蛋白的筛选与鉴定

Screening and identification of proteins interacting with cofilin CFL2 inmouse muscle fibers

筛选并鉴定小鼠腿肌组织中与丝切蛋白CFL2相互作用的蛋白质。通过免疫共沉淀技术分离小鼠腿肌组织中CFL2蛋白复合物,对获得的免疫共沉淀复合物进行SDS-PAGE分离,将分离出的差异条带用胰蛋白酶进行酶切,酶切后的肽段进行液相色谱分离,最后通过电喷雾质谱技术分析肽段组成并鉴定蛋白质,获得与CFL2互作的组织蛋白谱。通过Western blot检测,确定在天然状态下小鼠腿肌组织中CFL2与Ndufb7之间存在相互作用。首次利用免疫共沉淀联合质谱技术成功从小鼠腿肌组织中筛选并鉴定出与CFL2相互作用的蛋白Ndufb7,为进一步研究CFL2在骨骼肌肌纤维形成中的作用提供理论基础。

This study is aimed to screen and identify the proteins interacting with cofilin 2（ CFL2） from mouse skeletal muscle in leg. The CFL2 complex was extracted by co-immunoprecipitation,and separated by SDS-PAGE. The differential bands were cut by enzyme trypsin. The peptide segments were separated by liquid chromatography,and then analyzed and identified by electrospray ionization mass spectrometry. Tissue protein profiling interacting with CFL2 was obtained. The protein Ndufb7 which interacts with CFL2 from mouse skeletal muscle in leg was verified by co-immunoprecipitation,mass spectrometry and Western blot. This study provides a theoretical basis for the further study of CFL2 in the formation of skeletal muscle fibers.