Ⅰ类α-甘露糖苷酶诱导HIV-1包膜糖蛋白降解及机制研究  

Degradation of HIV-1 envelope glycoprotein induced by ClassⅠα-mannosidases and the potential mechanism

作　　者：刘笑辰 王绍清 王立平 吴桐[2] 于秀文 王俊苹 张晓杰 张英博 樊丽 李鸿梅 LIU Xiaochen;WANG Shaoqing;WANG Liping;WU Tong;YU Xiuwen;WANG Junping;ZHANG Xiaojie;ZHANG Yingbo;FAN Li;LI Hongmei(School of Pathology,Qiqihar Medical University,Qiqihar 161006,Heilongjiang,China;School of Psychiatry,Qiqihar Medical University,Qiqihar 161006,Heilongjiang,China;Research Institute of Medicine and Pharmacy,Qiqihar Medical University,Qiqihar 161006,Heilongjiang,China;School of Medical Technology,Zhang Zhou Health Vocational College/Collaborative Innovation Center for Translation Medical Testing and Application Technology,Zhangzhou 363000,Fujian,China)

机构地区：[1]齐齐哈尔医学院病理学院,黑龙江齐齐哈尔161006 [2]齐齐哈尔医学院精神卫生学院,黑龙江齐齐哈尔161006 [3]齐齐哈尔医学院医药科学研究院,黑龙江齐齐哈尔161006 [4]漳州卫生职业学院医学技术学院/转化医学检测应用技术协同创新中心,福建漳州363000

出　　处：《中国艾滋病性病》2024年第10期1007-1012,共6页Chinese Journal of Aids & STD

基　　金：黑龙江省自然科学基金项目(LH2020C108);漳州卫生职业学院引进高层次人才科研项目(BSKYQD-6);齐齐哈尔医学科学院临床科研基金项目(QMSI2019L-05)。

摘　　要：目的探究Ⅰ类α-甘露糖苷酶诱导HIV-1包膜糖蛋白(HIV-1 Env)的降解及其机制。方法将携带红色荧光表达Ⅰ类α-甘露糖苷酶的质粒与携带绿色荧光表达HIV-1 Env的质粒共转染293T细胞,Western Blot检测Ⅰ类α-甘露糖苷酶对HIV-1 Env表达水平的影响;激光共聚焦成像系统观察Ⅰ类α-甘露糖苷酶的亚细胞定位及Ⅰ类α-甘露糖苷酶与HIV-1 Env之间的相互作用;内质网相关蛋白质降解(ERAD)途径抑制剂探讨Ⅰ类α-甘露糖苷酶诱导HIV-1 Env降解的可能途径。结果Ⅰ类α-甘露糖苷酶可显著降低HIV-1 Env的表达水平,降低幅度达25%至68%;敲低Ⅰ类α-甘露糖苷酶的表达水平后,HIV-1 Env表达水平升高;ERAD抑制剂可显著升高HIV-1 Env的表达水平;激光共聚焦显微镜观察结果显示,Ⅰ类α-甘露糖苷酶与HIV-1 Env之间存在相互作用;激光共聚焦显微镜观察结果显示Ⅰ类α-甘露糖苷酶定位于内质网和高尔基体,与其行使生物学功能的部位相同。结论Ⅰ类α-甘露糖苷酶可能通过ERAD途径诱导HIV-1 Env降解。Objective To explore the degradation of the HIV-1 envelope glycoprotein(HIV-1 Env)induced by ClassⅠα-mannosidases and the potential mechanism.Methods Plasmids expressing ClassⅠα-mannosidases labeled with mCherry and plasmids expressing HIV-1 Env labeled with EGFP were co-transfected into 293T cells.The expression level of HIV-1 Env was detected by western blotting,and interaction between ClassⅠα-mannosidase and HIV-1 Env was observed by a laser confocal imaging system.Localization of ClassⅠα-mannosidases was also observed by the laser confocal imaging system,and an endoplasmic reticulum-associated degradation(ERAD)pathway inhibitor was used to explore the potential mechanism of the degradation of HIV-1 Env induced by ClassⅠα-mannosidases.Results ClassⅠα-mannosidases decreased the expression level of HIV-1 Env.significantly;the knock down of ClassⅠα-mannosidases increased the expression level of HIV-1 Env;and the ERAD pathway inhibitor increased the expression level of HIV-1 Env.Interactions between ClassⅠα-mannosidases and HIV-1 Env were observed with a laser confocal microscope,which showed that ClassⅠα-mannosidases were localized to the endoplasmic reticulum and Golgi complex,where they carried out their biological functions.Conclusions ClassⅠα-mannosidases probably induced the degradation of HIV-1 Env through the ERAD pathway.

关 键 词：Ⅰ类α-甘露糖苷酶 1型艾滋病病毒包膜糖蛋白 激光共聚焦 内质网相关蛋白质降解途径 

分 类 号：R512.91[医药卫生—内科学] R373.9[医药卫生—临床医学]