甲鱼蛋白源寡肽体外对微管蛋白聚合-解聚调控模式的影响  

Effects of Soft-shelled Turtle Protein-derived Oligopeptides on Tubulin Polymerization-Depolymerization Regulatory Mode in Vitro

作　　者：余俊莹 褚白怡 韩宇 赵雨欣 刘彩琴 王楠 YU Junying;CHU Baiyi;HAN Yu;ZHAO Yuxin;LIU Caiqin;WANG Nan(College of Biology and Environmental Engineering,Zhejiang Shuren University,Hangzhou 310015,China)

机构地区：[1]浙江树人学院生物与环境工程学院,浙江杭州310015

出　　处：《食品工业科技》2024年第16期85-93,共9页Science and Technology of Food Industry

基　　金：浙江省基础公益项目(LGN22C200024);国家级大学生创新创业训练计划(202211842009)。

摘　　要：为探讨甲鱼蛋白源寡肽对肿瘤细胞微管蛋白活性的调控机制,本研究以前期获得的甲鱼蛋白源三条寡肽DEADLLA(D-7-A)、EAGVNDW(E-7-W)、GSISSGQV(G-8-V)为研究对象,评价其对微管蛋白聚合-解聚的影响,解析其与微管蛋白的作用模式,阐释其诱导肿瘤细胞凋亡作用的机制。结果表明,3条寡肽与长春新碱相似,均为微管蛋白聚合的抑制剂,IC_(50)值分别为3.72、5.01、5.95μmol/L。分子对接结果显示,D-7-A与α-微管蛋白4X1I活性中心结合位点为Ser178和Thr179;E-7-W与活性中心结合位点为Gln15、Thr225、Tyr224、Tyr210、Arg214;G-8-V与活性中心结合位点为Gln11、Ser178、Asp329,与α-微管蛋白具有较强的作用力。细胞实验结果表明,D-7-A、E-7-W、G-8-V均对A549细胞具有抑制作用,且具有剂量依赖性,其IC_(50)值分别为2003±72、1877±102和1789±137μmol/L。G-8-V对A549细胞的抑制效果最强,且具有诱导A549细胞凋亡的作用,并随着药物处理时间的增加,凋亡率明显提高,其作用机制是通过影响α-微管蛋白的价键,抑制微管蛋白的聚合,诱导细胞凋亡。To examine the regulatory mechanisms of oligopeptides derived from soft-shelled turtle protein on tubulin activity in tumor cells,this study employed three specific oligopeptides D-7-A,E-7-W,and G-8-V obtained from soft-shelled turtle protein.The objective was to assess their impact on tubulin polymerization and depolymerization,investigate their interaction with tubulin,and elucidate the mechanisms induce apoptosis in tumor cells.The results showed that the three oligopeptides exhibited similarities to vincristine and acted as inhibitors of tubulin polymerization,their respective IC_(50)values was 3.72,5.01,and 5.95µmol/L.Furthermore,the molecular docking analysis revealed that D-7-A interacted with the active center ofα-tubulin 4X1I,specifically binding to Ser178 and Thr179.The active center of E-7-W was found to be bound by Gln15,Thr225,Tyr224,Tyr210 and Arg214.Similarly,G-8-V was observed to bind to Gln11,Ser178 and Asp329 at the active center,exerting a significant impact onα-tubulin.The results of cell experiments showed that D-7-A,E-7-W and G-8-V had dose-dependent inhibitory effects on A549 cells,with IC_(50)values of 2003±72,1877±102 and 1789±137μmol/L,respectively.The G-8-V exhibited the most potent inhibitory effect on A549 cells,leading to the induction of apoptosis.Moreover,the apoptosis rate demonstrated a significant increase with prolonged drug treatment time.The underlying mechanism of action involved the inhibition of tubulin polymerization and the induction of apoptosis by perturbing the valence bond ofα-tubulin.

关 键 词：甲鱼蛋白源寡肽 微管蛋白聚合 抑制作用 细胞凋亡 

分 类 号：TS253.9[轻工技术与工程—农产品加工及贮藏工程]