去棕榈酰化修饰下G蛋白偶联受体激酶6核定位信号鉴定  

Identification of Nuclear Localization Sequence of G Protein-Coupled Receptor Kinase 6 under Depalmitoylation

作　　者：黄希仕 李瑞瑞 黄岚珍[1,2] 阳晶 吴群英[1,3] 蒋晓山 Huang Xishi;Li Ruirui;Huang Lanzhen;Yang Jing;Wu Qunying;Jiang Xiaoshan(Cell Signaling Laborary, Guilin Medical University, Guilin 541004, China;Center for Science Research, Guilin Medical University, Guilin 541004, China;Department of Genetics and Cell Biology, GuiIin Medical University, Guilin 541004, China;Graduate College, Guilin Medical University, Guilin 541004, China)

机构地区：[1]桂林医学院细胞信号转导实验室,桂林541004 [2]桂林医学院科学实验中心,桂林541004 [3]桂林医学院细胞与遗传教研室,桂林541004 [4]桂林医学院研究生学院,桂林541004

出　　处：《中国细胞生物学学报》2018年第6期955-961,共7页Chinese Journal of Cell Biology

基　　金：国家自然科学基金项目(批准号:31460306;31660327);广西自然科学基金项目(批准号:2014GXNSFAA118158);广西研究生教育创新计划项目(批准号:YCSW2017207)资助的课题~~

摘　　要：G蛋白偶联受体激酶6(G protein-coupled receptor kinase 6,GRK6)依赖去棕榈酰化修饰及特定核定位序列(nuclear localization sequence,NLS)实现胞核定位,但NLS的关键基团及其对激酶活性的影响尚不清楚。该研究首先通过构建一系列缺失突变子,初筛去棕榈酰化条件下GRK6的NLS结构域;然后采用点突变技术进一步确定NLS结构域中Lys(K)^(389)、Lys(K)^(390)、Lys(K)^(3913)个关键基团;最后通过检测内源性毒蕈碱M3受体介导的细胞内钙流,证实NLS突变子对M3受体介导的细胞内钙流信号的抑制作用无明显影响。该研究为进一步揭示GRK6胞核转运机制及其功能提供了有价值的信息。G protein-coupled receptor kinase 6（GRK6） is capable of achieving nuclear translocation through depalmitoylation. Although a nuclear localization sequence（NLS） region has reported, the key residue（s） in the NLS domain as well as its effect on the kinase activity are not yet clear. In this study, we first verified a predicted NLS domain through screening a series of GRK6 deletion mutants under the condition of depalmitoylation. Then, we further identified three residues, Lys（K）（389）, Lys（K）（390） and Lys（K）（391）, which respectively played essential role in the nuclear translocalization of GRK6 by using a point mutation technology. Finally, the biological function of the mutants in the M3 receptor signaling, comparing to GRK6 WT, was analyzed using a calcium mobilization assay and found no significant effect on their kinase activities. This study provides valuable information to further study the nuclear translocation mechanism and function of GRK6.

关 键 词：GRK6 去棕榈酰化 核定位信号 G蛋白偶联受体激酶 

分 类 号：Q51[生物学—生物化学]