猪精浆液中磷脂酰乙醇胺结合蛋白4的纯化及其一级结构分析  

Purification and primary structure analysis of phosphatidylethanolamine-binding protein 4 from porcine seminal plasma

作　　者：安丽萍[1] 杜瑞丽[1] 徐广宇[1] 韩笑[1] 詹金卓[1] 前田利长 大久保岩男 杜培革[1] 

机构地区：[1]北华大学药学院微生物与生化药学教研室,吉林吉林132013 [2]日本滋贺医科大学

出　　处：《吉林大学学报（医学版）》2010年第6期1196-1200,共5页Journal of Jilin University：Medicine Edition

基　　金：教育部留学回国人员科研启动基金资助课题(教外司留[2004]176号)

摘　　要：目的:从猪精浆中纯化磷脂酰乙醇胺结合蛋白家族的一个新成员——猪磷脂酰乙醇胺结合蛋白4(sus PEBP4),为进一步研究其功能奠定理论基础。方法:利用饱和硫酸铵盐析、Q-Sepharose、Matrex Gel RedA、SP-XL-Sepharose和FPLC Mono-S(A)柱层析从500mL猪精浆中纯化susPEBP4,自动氨基酸测序仪进行N-末端氨基酸测序及内部序列分析。生物信息学方法分析其与人、牛和狗的序列同源性。结果:经SDS-PAGE电泳分离鉴定得到了相对分子质量为25000的单一区带的sus PEBP4。生物信息学方法分析显示,其与人、牛和狗之间氨基酸序列同源性为75.2%～81.8%之间。结论:从猪精浆中制备了电泳纯的PEBPs家族的新成员sus PEBP4,明确了其一级结构,生物信息学分析显示其与其他生物体氨基酸同源性高。Objective To purify the phosphatidylethanolamine-binding protein 4（PEBP4）,a new member of the phosphatidylethanolamine-binding protein family from porcine seminal plasma and lay the foundation for further investigation of its function.Methods Sus PEBP4was purified from approximately 500mL porcine seminal plasma by ammonium sulfate fractionation, Q-Sepharose column chromatography, matrex Gel Red A column chromatography,SP-XL-Sepharose column chromatography and Mono-S （A）column chromatography（in FPLC）. The amino acid sequence of the purified sus PEBP4was identified by automated N-terminal sequencing of the intact protein.The amino acid sequences of sus PEBP4were compared with those of human,bovine,dog PEBP4by bioinformatics.Results The molecular mass of the purified protein was calculated to be 25 000by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence of beta-mercaptoethanol.The overall homology of amino acid sequence between sus,human,bovine and dog was from 75.2to 81.8%.Conclusion Sus PEBP4is purified by the optimal methods from porcine seminal plasma.Its primary structure is defined.Sus PEBP4has high homology with other organisms.

关 键 词：磷脂酰乙醇胺结合蛋白4 纯化 猪精浆 氨基酸序列 

分 类 号：Q7[生物学—分子生物学]