人WASH468和WASH465蛋白质特性辨析  

Comparison of Protein Characteristics of Human WASH468 and WASH465

作　　者：郑皑雪 张鲁平 汤拓 李萍 洪鲜 邓志会 王涛 ZHENG Aixue;ZHANG Luping;TANG Tuo;LI Ping;HONG Xian;DENG Zhihui;WANG Tao(Laboratory of Protein Structure and Function,Research Institute of Medicine and Pharmacy,Qiqihar Medical University,Qiqihar,Heilongjiang,161000,China;College of Medical Technology,Qiqihar Medical University,Qiqihar,Heilongjiang,161000,China)

机构地区：[1]齐齐哈尔医学院医药科学研究院,蛋白质结构与功能研究室,黑龙江省齐齐哈尔市161000 [2]齐齐哈尔医学院医学技术学院,黑龙江省齐齐哈尔市161000

出　　处：《医学分子生物学杂志》2024年第2期93-99,共7页Journal of Medical Molecular Biology

基　　金：黑龙江省自然科学基金优秀青年项目(No.YQ2022C036);国家自然科学基金青年项目(No.31800648);黑龙江省教育厅优秀创新团队项目(No.2020-KYYWF-0001);齐齐哈尔医学院研究生创新基金项目(No.QYYCX2022-06)。

摘　　要：目的探讨两种较为公认但序列不同的Wiskott-Aldrich综合征蛋白和富含脯氨酸同源物(Wiskott-Aldrich syndrome protein and SCAR homolog,WASH)的差异。方法使用免疫荧光、免疫共沉淀和激光微辐射等实验分析两种WASH在定位模式、与FAM21或Ku蛋白的相互作用、向DNA损伤位点募集速率和蛋白质稳定性方面的差异;比较多种生物中的WASH蛋白质序列和检测多种在生物和医学研究中常用的细胞中的WASH编码序列,分析两种人WASH序列的普遍性和保守性。结果两种WASH展现出类似的内体(endosome)定位模式。WASH468表现出与FAM21更强的相互作用,并且WASH468展现出更强的稳定性。WASH465表现出与Ku蛋白更强的相互作用,并且WASH465展现出向DNA损伤位点更强的募集。多种生物中的WASH序列与人WASH468序列的一致性明显高于WASH465,并且多种在生物和医学研究中常用的细胞中的WASH氨基酸序列均与WASH468一致。结论WASH468和WASH465的生物学特性存在差异,WASH468序列的普遍性和保守性明显高于WASH465,因此WASH468是更保守的人WASH序列。Objective To investigate the differences between two well recognized Wiskott-Aldrich syndrome protein and SCAR homolog(WASH)with different sequences.Methods Immunofluorescence,immunoprecipitation,and laser micro-irradiation were used to analyze the differences between WASH468 and WASH465 in localization patterns,the interaction with FAM21 or Ku proteins,the recruitment to DNA damage sites and the protein stabilities.To analyze the universality and conservation of the two types of WASH sequences,the WASH protein sequences in various organisms were compared,and the WASH coding sequences in various cell types commonly used for biological and medical research were detected.Results Both WASH exhibited a similar way in localizing to endosome.WASH468 exhibited a stronger interaction with FAM21,and was more stable than WASH465.WASH465 exhibited a stronger interaction with Ku protein,and the recruitment of WASH465 to DNA damage sites was more robust than that of WASH468.The degree of similarity observed between the sequence of WASH468 in humans and the sequences of WASH in other organisms was significantly greater than that observed with WASH465.Moreover,the coding sequences of WASH in various cell types,which are commonly utilized in biological and medical research,exhibited concordance with the sequence of WASH468.Conclusion There are differences in the biological characteristics between WASH468 and WASH465,and WASH468 is significantly more universal and conserved than WASH465.Therefore,WASH468 is the conserved human WASH sequence.

关 键 词：Wiskott-Aldrich综合征蛋白和富含脯氨酸同源物 蛋白序列 保守性 相互作用 DNA修复 

分 类 号：Q71[生物学—分子生物学] R34[医药卫生—基础医学]