项东鑫, 郑寓, 周诗航, 陆奕, 王艳伟. 氯化钾及聚乙二醇对聚脯氨酸-精氨酸液-液相分离的协同影响[J]. 分析测试技术与仪器, 2023, 29(1): 1-6. DOI: 10.16495/j.1006-3757.2023.01.001
引用本文: 项东鑫, 郑寓, 周诗航, 陆奕, 王艳伟. 氯化钾及聚乙二醇对聚脯氨酸-精氨酸液-液相分离的协同影响[J]. 分析测试技术与仪器, 2023, 29(1): 1-6. DOI: 10.16495/j.1006-3757.2023.01.001
XIANG Dongxin, ZHENG Yu, ZHOU Shihang, LU Yi, WANG Yanwei. Cooperative Effect of KCl and Polyethylene Glycol on Liquid-Liquid Phase Separation of Polyproline-Arginine[J]. Analysis and Testing Technology and Instruments, 2023, 29(1): 1-6. DOI: 10.16495/j.1006-3757.2023.01.001
Citation: XIANG Dongxin, ZHENG Yu, ZHOU Shihang, LU Yi, WANG Yanwei. Cooperative Effect of KCl and Polyethylene Glycol on Liquid-Liquid Phase Separation of Polyproline-Arginine[J]. Analysis and Testing Technology and Instruments, 2023, 29(1): 1-6. DOI: 10.16495/j.1006-3757.2023.01.001

氯化钾及聚乙二醇对聚脯氨酸-精氨酸液-液相分离的协同影响

Cooperative Effect of KCl and Polyethylene Glycol on Liquid-Liquid Phase Separation of Polyproline-Arginine

  • 摘要: 由脯氨酸(Proline,P)和精氨酸(Arginine,R)构成的双重复肽是人体内C9ORF72基因异常扩增产生的富含毒性的多肽,其异常扩增及液-液相分离(liquid-liquid phase separation,LLPS)行为是神经退行性疾病发病的重要原因,因此研究多肽的调控行为可以为治疗相关疾病提供帮助. 主要利用光学显微成像以及紫外分光光度计技术,研究35个脯氨酸-精氨酸重复多肽(PR35)的LLPS现象及其调控,并对机制进行分析. 发现PR35的LLPS的发生依赖溶液中氯化钾的浓度,当氯化钾浓度低于2 700 mmol/L时,LLPS不会出现,而其浓度高于2 700 mmol/L时,LLPS会发生. 同时在此临界浓度(2 700 mmol/L)溶液中,加入不同质量分数的分子拥挤剂聚乙二醇(PEG1000)后,发现PEG对LLPS具有促进作用,并且随着PEG质量分数升高,促进作用增强. 同时通过紫外分光光度计对吸光度进行了分析,得到的结论与显微镜的观察结果一致.

     

    Abstract: The double complex peptide composed of proline (Proline, P) and arginine (Arginine, R) is a toxicity-rich polypeptide produced by the abnormal amplification of C9ORF72 gene in the human body. The abnormal amplification and liquid-liquid phase separation (LLPS) behavior of the peptide are important causes of neurodegenerative diseases, so to study the regulatory behavior of the peptide is helpful to the therapeutic treatment of related diseases. In the paper, optical microscopic imaging and ultraviolet spectrophotometric technique were used to study the LLPS phenomenon and the regulation of related 35 proline-arginine repeat peptides (PR35), and the regulation mechanism was analyzed. It was found that the occurrence of LLPS in PR35 depended on the concentration of potassium chloride in the solution. LLPS did not take place when the concentration of KCl was lower than 2 700 mmol/L, but LLPS took place when the concentration was higher than 2 700 mmol/L. In this critical concentration (2 700 mmol/L) of the KCl solution, after adding different mass fractions of the molecular crowding agent polyethylene glycol (PEG1000), it was found that PEG had a promoting effect on LLPS, and the promoting effect increased with the increase of mass fraction of PEG concentration. At the same time, the conclusion of ultraviolet spectrophotometric absorbance analysis was consistent with the results of microscopic survey.

     

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