高等学校化学学报 ›› 2016, Vol. 37 ›› Issue (11): 1987-1992.doi: 10.7503/cjcu20160417

• 有机化学 • 上一篇    下一篇

K33位乙酰化修饰SUMO蛋白的化学合成

王业海1, 孔一夫1, 陈晨晨2, 李宜明1()   

  1. 1. 合肥工业大学生物与医学工程学院, 合肥 230009
    2. 中国科学院强磁场科学中心, 合肥 230031
  • 收稿日期:2016-06-12 出版日期:2016-11-10 发布日期:2016-10-19
  • 作者简介:联系人简介: 李宜明, 男, 博士, 副教授, 主要从事翻译后修饰蛋白的化学合成、 结构及功能方面的研究. E-mails: lym2007@mail.ustc.edu.cn; ymli@hfut.edu.cn
  • 基金资助:
    国家自然科学基金(批准号: 21372058, 21572043)资助

Chemical Synthesis of K33 Acetylated SUMO Protein

WANG Yehai1, KONG Yifu1, CHEN Chenchen2, LI Yiming1,*()   

  1. 1.School of Biological and Medical Engineering, Hefei University of Technology, Hefei 230009, China
    2. High Magnetic Field Laboratory of the Chinese Academy of Sciences, Hefei 230031, China
  • Received:2016-06-12 Online:2016-11-10 Published:2016-10-19
  • Contact: LI Yiming E-mail:ymli@hfut.edu.cn
  • Supported by:
    † Supported by the National Natural Science Foundation of China(Nos.21372058, 21572043)

摘要:

采用高温辅助固相合成技术及基于多肽酰肼的自然化学连接技术, 高效获得了小泛素相关修饰物(SUMO)蛋白及33位赖氨酸(K33)乙酰化SUMO蛋白. 聚丙烯酰胺凝胶电泳(SDS-PAGE)和圆二色光谱分析结果表明, 与生物表达蛋白相比, 化学合成蛋白具有较好的纯度和类似的二级结构.

关键词: 小泛素相关修饰物, 乙酰化, 高温辅助固相合成, 多肽酰肼, 自然化学连接

Abstract:

Sumoylated protein was found to reduce the protein binding force with the SUMO-interaction motif(SIM) of effector protein when K33 of SUMO was acetylated. To further study the structures and mechanism, a large amount of uniformity acetylated SUMO protein was greatly needed. In this work, SUMO protein and K33 acetylated SUMO protein was efficiently obtained using high temperature assisted solid-phase peptide synthesis(SPPS) technology combined with peptide hydrazide ligation. SUMO and K33 acetylated SUMO was identified with corrected molecular weight by the SDS-PAGE. It was also confirmed that the chemically synthe-tic protein and biologically expressed SUMO2 own similar homogeneity and secondary structure by the CD analysis. These results provided a foundation for the follow-up study of K33 acetylated SUMO protein.

Key words: Small ubquitin-related modifier, Acetylation, High temperature assisted solid-phase peptide synthesis, Peptide hydrazide, Native chemical ligation

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