摘要
细胞内大量的无膜室(统称为生物大分子凝聚体)是通过液-液相分离现象(LLPS)组装起来的。相分离凝聚体参与多种生物学活动,而异常的相分离可能会导致大量复杂的人类疾病,如神经退行性疾病和癌症等。因此,阐明多肽和DNA的液-液相分离现象背后隐藏的机制可能对理解和治疗一些严重的人类疾病提供指导。该研究主要探索一系列含有重复赖氨酸和组氨酸(KH)序列的多肽(Poly KH)相关LLPS的调控,研究发现多肽KH-30与DNA通过静电相互作用产生LLPS现象,室温下DNA调控相分离的质量浓度范围为6~14μg/μL,质量浓度进一步增大LLPS现象则会消失。在DNA和KH-30多肽的混合溶液中加入PEG-1000,发现其分子拥挤效应促进了小液滴凝聚为大液滴,随着PEG-1000浓度增加,分子拥挤效应越明显,越有利于LLPS现象的发生。此外,温度是体外调控LLPS现象的便捷手段,本研究发现KH-30表现出熵驱动的LLPS行为,即温度的升高抑制了KH-30多肽与DNA的LLPS现象。
A large number of membrane-free compartments within cells(collectively known as bio macromolecular condensates)are assembled through the phenomenon of liquid-liquid phase separation(LLPS).Phase-separated condensates participate in various biological activities,and abnormal phase separation may lead to a large number of complex human diseases,such as neurodegenerative diseases and cancer.Therefore,elucidating the mechanisms underlying the liquid-liquid phase separation of peptides and DNA may provide guidance for understanding and treating some severe human diseases.In this study,the regulation of LLPS was mainly explored related to a series of peptides(Poly KH)containing repeated lysine and histidine(KH)sequences.It was found that the KH-30 peptide produced LLPS with DNA through electrostatic interactions.At room temperature,the range of DNA-regulated phase separation was 6~14μg/μL,and further increases in concentration led to the disappearance of the LLPS phenomenon.When PEG-1000 was added to a mixed solution of DNA and KH-30 peptides,its molecular crowding effect promoted the aggregation of small droplets into large droplets.As the concentration of PEG-1000 increased,the molecular crowding effect became more apparent,favoring the occurrence of the LLPS phenomenon.In addition,temperature is a convenient means to regulate LLPS in vitro.This study found that KH-30 exhibited entropy-driven LLPS behavior,meaning that an increase in temperature restrained the LLPS phenomenon between the KH-30 peptide and DNA.
作者
陈思远
严欢
何雪童
杨光参
CHEN Siyuan;YAN Huan;HE Xuetong;YANG Guangcan(School of Mathematics and Physics,Wenzhou University,Wenzhou 325035,China)
出处
《齐鲁工业大学学报》
2025年第2期53-60,共8页
Journal of Qilu University of Technology
基金
国家自然科学基金(12074289、11574232)
浙江省自然科学基金面上项目(Y23A040004)
浙江省高等教育“十四五”教学改革项目(jg20220509)
浙江省研究生教育学会科研项目(2023-016)
浙江省新苗(2023R451012)
国家级创新创业计划(JWXC2022070)。
