摘要
为揭示HMGB4在精子发生过程中所起的作用,采用免疫共沉淀技术富集NTera-2细胞中的HMGB4结合蛋白.SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)对免疫沉淀复合物进行分离,从胶中切取HMGB4结合蛋白条带,胶内酶解后进行电喷雾串联质谱(LC-ESI-MS/MS)分析,得到相应的肽序列标签(peptide sequence tags,PST).通过搜索数据库在NTera-2细胞系中鉴定到5个HMGB4相互作用蛋白,分别是Pdha1、Keratin 6B、Keratin 5、Actb Actin cytoplasmic 1、RPL18.为进一步研究HMGB4在精子发生机制中的生理功能提供了基础.
In order to reveal the role of HMGB4 in the process of spermatogenesis, the binding proteins of HMGB4 were enriched in NTera-2 ceils using co-immunoprecipitation assay. The immunoprecipitates were separa- ted on 10% SDS-polyacrylamide gel, then the binding proteins were cut from the gel and analyzed by LC-ECI-MS/ MS after the enzymolysis. The corresponding peptide sequence tags（PST） and identified 5 interacting proteins of HMGB4 were obtained in the NTera-2 cells using the database. These interacting proteins, such as Pdhal, Keratin 6B, Keratin 5, Actb Actin cytoplasmic 1, RPL18, established foundation for the further study on physiological functions of HMGB4 in the mechanism of spermatogenesis.
出处
《湖南师范大学自然科学学报》
CAS
北大核心
2012年第2期71-75,共5页
Journal of Natural Science of Hunan Normal University
基金
国家自然科学基金资助项目(81071696)
湖南省研究生科研创新资助项目(CX2011B214)
教育部留学回国人员科研启动基金资助项目((2010)609号)
长沙市科技攻关资金专项资助项目(K1109006-31)
