Bacillus subtilis转录因子CtsR蛋白中clpC操纵子结合区域的NMR研究

doi:10.11938/cjmr20202854

波谱学杂志 ›› 2021, Vol. 38 ›› Issue (2): 155-163.doi: 10.11938/cjmr20202854

Bacillus subtilis转录因子CtsR蛋白中clpC操纵子结合区域的NMR研究

陈晓雯¹,黄碧玲¹,黄少华^1,*(),赵玉芬^1,^2,^3,*()

1. 宁波大学新药技术研究院, 浙江宁波 315211
2. 厦门大学化学化工学院, 福建省化学生物学重点实验室, 福建厦门 361005
3. 清华大学化学系, 生命有机磷化学与化学生物学重点实验室, 北京 100084

收稿日期:2020-09-21 出版日期:2021-06-05 发布日期:2020-11-20
通讯作者: 黄少华,赵玉芬 E-mail:huangshaohua@nbu.edu.cn;zhaoyufen@nbu.edu.cn
基金资助:
国家自然科学基金资助项目(91856126);福建省化学生物学重点实验室（厦门大学）开放基金资助项目(2021001)

An NMR Study on the clpC Operon Binding Region of Transcription Factor CtsR from Bacillus subtilis

Xiao-wen CHEN¹,Bi-ling HUANG¹,Shao-hua HUANG^1,*(),Yu-fen ZHAO^1,^2,^3,*()

1. Institute of Drug Discovery Technology, Ningbo University, Ningbo 315211, China
2. Department of Chemical Biology, College of Chemistry and Chemical Engineering, and the Key Laboratory for Chemical Biology of Fujian Province, Xiamen University, Xiamen 361005, China
3. Key Lab of Bioorganic Phosphorus Chemistry & Chemical Biology, Department of Chemistry, Tsinghua University, Beijing 100084, China

Received:2020-09-21 Online:2021-06-05 Published:2020-11-20
Contact: Shao-hua HUANG,Yu-fen ZHAO E-mail:huangshaohua@nbu.edu.cn;zhaoyufen@nbu.edu.cn

摘要/Abstract

摘要：

革兰氏阳性菌枯草芽孢杆菌（Bacillus subtilis）响应热刺激时，精氨酸激酶McsB磷酸化转录因子CtsR蛋白中clpC操纵子结合区域的Arg62（R）位点，使得CtsR与clpC操纵子解离，从而启动clpC相关基因的转录过程，以表达细菌应对热刺激所需的蛋白.本文以CtsR蛋白中结合clpC操纵子的区域（KRGGGG）为研究对象，通过对¹H NMR、¹H-¹H COSY、¹H-¹H TOCSY、¹H-¹⁵N HSQC和¹H-¹³C HSQC等谱图的综合分析，对其¹H、¹³C以及¹⁵N的化学位移进行归属，为该片段与clpC操纵子相互作用，以及精氨酸磷酸化调控机制的研究提供基础.

关键词: 转录因子, 核磁共振, 化学位移, 溶液结构

Abstract:

When Gram-positive bacterium Bacillus subtilis responds to heat-shock, protein arginine kinase McsB phosphorylates Arg62 (R) in the clpC operon binding region of transcription factor CtsR, leading to dissociation of CtsR and clpC operon and initiation of stress-genes transcription for the ensuing expression of heat-shock proteins. This work investigated the clpC operon binding region (KRGGGG) of CtsR with NMR spectra (i.e., ¹H NMR, ¹H-¹H COSY, ¹H-¹H TOCSY, ¹H-¹⁵N HSQC and ¹H-¹³C HSQC). The ¹H, ¹³C and ¹⁵N chemical shifts of the protein segment were assigned. The interaction between CtsR and clpC operon and the regulatory mechanism of arginine phosphorylation were analyzed.

Key words: transcription factor, nuclear magnetic resonance (NMR), chemical shifts, structure in solution

中图分类号:

O482.53

陈晓雯,黄碧玲,黄少华,赵玉芬. Bacillus subtilis转录因子CtsR蛋白中clpC操纵子结合区域的NMR研究[J]. 波谱学杂志, 2021, 38(2): 155-163.

Xiao-wen CHEN,Bi-ling HUANG,Shao-hua HUANG,Yu-fen ZHAO. An NMR Study on the clpC Operon Binding Region of Transcription Factor CtsR from Bacillus subtilis[J]. Chinese Journal of Magnetic Resonance, 2021, 38(2): 155-163.

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Position	δ_H/ppm	δ_N/ppm	δ_C/ppm
K1-α	4.07	/	52.90
K1-β	1.92	/	30.65
K1-γ	1.45	/	21.04
K1-δ	1.70	/	26.47
K1-ε	3.01	/	39.35
R2-NH-α	8.83	124.60	/
R2-α	4.38	/	/
R2-β	1.83	/	28.12
R2-γ	1.69	/	24.43
R2-δ	3.22	/	40.64
R2-NH-ε	7.18	84.72	/
G3-NH-α	8.66	111.72	/
G3-α	3.98	/	42.54
G4-NH-α	8.37/8.38	109.16/108.91	/
G4-α	4.01/4.02	/	42.54
G5-NH-α	8.37/8.38	109.16/108.91	/
G5-α	4.01/4.02	/	42.54
G6-NH-α	8.08	114.16	/
G6-α	3.83	/	43.00

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Bacillus subtilis转录因子CtsR蛋白中clpC操纵子结合区域的NMR研究

An NMR Study on the clpC Operon Binding Region of Transcription Factor CtsR from Bacillus subtilis

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