K33位乙酰化修饰SUMO蛋白的化学合成

doi:10.7503/cjcu20160417

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

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. 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 Yiming^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/Abstract

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.

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

中图分类号:

O629

TrendMD:

王业海, 孔一夫, 陈晨晨, 李宜明. K33位乙酰化修饰SUMO蛋白的化学合成. 高等学校化学学报, 2016, 37(11): 1987.

WANG Yehai, KONG Yifu, CHEN Chenchen, LI Yiming. Chemical Synthesis of K33 Acetylated SUMO Protein^†. Chem. J. Chinese Universities, 2016, 37(11): 1987.

图/表 6

Scheme 1 Amino acid sequence of SUMO2(A) and the synthesis strategy of SUMO2 and K33-Ac-SUMO2(B)

Fig.1 HPLC trace(A, B, C) and MS(D, E, F) of SUMO2-1(A, D), K33-Ac-SUMO2-1(B, E) and SUMO2-2(C, F)

Fig.2 Ligation trace of SUMO2-1(A) and SUMO2-2(B), and MS(C) of SUMO2

Fig.3 Ligation trace of K33-Ac-SUMO2-1(A) and SUMO2-2(B), and MS(C) of K33-Ac-SUMO2

Fig.4 SDS-PAGE of expressed SUMO2(1), synthesized SUMO2(2) and synthesized K33-Ac-SUMO2(3)

Fig.5 CD spectra of expressed SUMO2, synthesized SUMO2 and synthesized K33-Ac-SUMO2

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K33位乙酰化修饰SUMO蛋白的化学合成

Chemical Synthesis of K33 Acetylated SUMO Protein^†

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K33位乙酰化修饰SUMO蛋白的化学合成

Chemical Synthesis of K33 Acetylated SUMO Protein†

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Chemical Synthesis of K33 Acetylated SUMO Protein^†