To refine the signal transduction network mediated by the protein tyrosine kinase LYN and clarify its regulatory mechanism in allergic diseases, this study used a yeast two-hybrid system to screen target proteins that specifically interact with LYN. Self-activation activity and cytotoxicity of the bait protein were verified using haploid and diploid mating assays. Primary positive clones were subjected to sequencing alignment and bioinformatics analysis, and protein-protein interactions were further validated by yeast reversion experiments. A total of 53 potential interacting proteins were obtained in the primary screening. After removing invalid sequences including frameshift mutations, non-coding region translations, and coding region deletions, 10 candidate interacting proteins were retained. Combining bioinformatics analysis and yeast reversion experiments validation, the specific interaction between LYN and polyubiquitin-B precursor(UBB)was finally confirmed. This study provides an experimental basis for- 引用格式:冯忠居,于明威,张聪,等.冲刷场地地震波形对大直径变截面单桩时程响应的影响［J］.河北大学学报(自然科学版),2026,46(4):337-348.引用格式:龙通云,杨艳蕾,齐梦楠,等.蛋白酪氨酸激酶LYN互作蛋白筛选［J］.河北大学学报(自然科学版),2026,46(4):374-382.DOI:10.3969/j.issn.1000-1565.2026.04.005蛋白酪氨酸激酶LYN互作蛋白筛选龙通云,杨艳蕾,齐梦楠,张艳芬,刘中成(河北大学 药学院,河北省药物质量分析控制重点实验室,河北 保定 071002)摘 要:为完善蛋白酪氨酸激酶LYN介导的信号转导网络,明确其在变态反应性疾病中的调控机制,采用酵母双杂交系统筛选与LYN存在特异性相互作用的靶蛋白.通过单倍体与二倍体接合实验验证诱饵蛋白的自激活活性与细胞毒性,对初筛获得的阳性克隆进行测序比对和生物信息学分析,并通过酵母回转实验验证蛋白间互作关系.经初筛获得53个潜在互作蛋白,剔除移码突变、非编码区翻译及编码区翻译缺失等无效序列后,得到10个候选互作蛋白.结合生物信息学预测与回转实验验证,最终证实LYN与多聚泛素B前体(polyubiquitin-B precursor,UBB)存在特异性相互作用.本研究为深入阐明LYN在变态反应性疾病中的调控机制提供了实验依据,也为相关疾病治疗提供了新的潜在靶点.关键词:LYN;酵母文库;酵母双杂交;互作蛋白;生物信息学分析中图分类号:R593.1 文献标志码:A 文章编号:1000-1565(2026)04-0374-09DOI:10.3969/j.issn.1000-1565.2026.04.005Screening of proteins interacting with protein tyrosine kinase LYNLONG Tongyun, YANG Yanlei, QI Mengnan, ZHANG Yanfen, LIU Zhongcheng(Key Laboratory of Pharmaceutical Quality Control of Hebei Province, College of Pharmaceutical Sciences, Hebei University, Baoding 071002, China)Abstract: To refine the signal transduction network mediated by the protein tyrosine kinase LYN and clarify its regulatory mechanism in allergic diseases, this study used a yeast two-hybrid system to screen target proteins that specifically interact with LYN. Self-activation activity and cytotoxicity of the bait protein were verified using haploid and diploid mating assays. Primary positive clones were subjected to sequencing alignment and bioinformatics analysis, and protein-protein interactions were further validated by yeast reversion experiments. A total of 53 potential interacting proteins were obtained in the primary screening. After removing invalid sequences including frameshift mutations, non-coding region translations, and coding region deletions, 10 candidate interacting proteins were retained. Combining bioinformatics analysis and yeast reversion experiments validation, the specific interaction between LYN and polyubiquitin-B precursor(UBB)was finally confirmed. This study provides an experimental basis for- 收稿日期:2026-02-13;修回日期:2026-04-07 基金项目:河北省自然科学基金项目(H2022201035;H2023201026) 第一作者:龙通云(1999—), 女, 河北大学在读硕士研究生, 主要从事基因工程制药和分子药理学方向研究.E-mail:2742486460@qq.com 通信作者:刘中成(1979—), 男, 河北大学教授, 主要从事基因工程制药和分子药理学方向研究.E-mail:liuzc@hbu.edu.cn 第4期龙通云等:蛋白酪氨酸激酶LYN互作蛋白筛选河北大学学报(自然科学版) 第46卷further elucidating the regulatory mechanism of LYN in allergic diseases and offers novel potential targets for the treatment of related disorders.
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