• 中国中文核心期刊
  • 中国科学引文数据库核心期刊
  • 中国科技核心期刊
  • 中国高校百佳科技期刊
高级检索

N-酰基葡糖胺衍生物对GABA转运体功能的影响

胡静, 樊华, 尹健

胡静, 樊华, 尹健. N-酰基葡糖胺衍生物对GABA转运体功能的影响[J]. 中国药科大学学报, 2016, 47(2): 228-234. DOI: 10.11665/j.issn.1000-5048.20160217
引用本文: 胡静, 樊华, 尹健. N-酰基葡糖胺衍生物对GABA转运体功能的影响[J]. 中国药科大学学报, 2016, 47(2): 228-234. DOI: 10.11665/j.issn.1000-5048.20160217
shu
HU Jing, FAN Hua, YIN Jian. Effects of N-acyl glucosamines on GABA uptake activity of GABA transporter 1[J]. Journal of China Pharmaceutical University, 2016, 47(2): 228-234. DOI: 10.11665/j.issn.1000-5048.20160217
Citation: HU Jing, FAN Hua, YIN Jian. Effects of N-acyl glucosamines on GABA uptake activity of GABA transporter 1[J]. Journal of China Pharmaceutical University, 2016, 47(2): 228-234. DOI: 10.11665/j.issn.1000-5048.20160217
shu

N-酰基葡糖胺衍生物对GABA转运体功能的影响

  • 1.

    江南大学无锡医学院

  • 2.

    柏林自由大学夏洛特医学院

  • 3.

    江南大学生物工程学院糖化学与生物技术教育部重点实验室

基金项目: 江苏省自然基金资助项目(No.BK20150140,No.BK20140154)

Effects of N-acyl glucosamines on GABA uptake activity of GABA transporter 1

摘要

    摘要
  • 摘要: 为筛选GAT蛋白活性抑制剂，以稳定表达GAT1蛋白的人胚胎肾细胞(HEK-293)为模型，利用GABA摄取定量测试测定N-酰基葡糖胺衍生物对该蛋白功能的影响。同时，由于GAT1蛋白的功能与其N-糖链结构尤其是末端唾液酸密切相关，再通过定量该蛋白糖链末端唾液酸的表达分析衍生物对蛋白糖链修饰的作用。结果显示，10 mmol/L 3-O-甲基-N-乙酰基葡糖胺(3-O-Met-GlcNAc)通过抑制GAT1蛋白的唾液酸化(降至66.8%)将GABA摄取活性减弱到到53%(P<；0.01)；而10 mmol/L N-丙酰基葡糖胺(GlcNProp)，N-环丙甲酰基葡糖胺(GlcNCyclo)，N-己酰基葡糖胺(GlcNHex)，N-乙酰氨基乙酰基葡糖(GlcNAc-acetamido)则通过抑制蛋白糖链末端修饰分别将GABA摄取活性减弱到54%、63%、63%和67%(P<；0.01)。因此，N-酰基葡糖胺衍生物可用于进一步筛选研究GAT蛋白活性抑制剂或唾液酸生物合成抑制剂。
    Abstract: In order to screen the inhibitors of GAT1 protein, the effects of synthetic N-acylglucosamines on the GABA uptake activity of GAT1 were examined in the HEK-293 cell model stably expressing GAT1 by performing GABA uptake assay. And since the N-linked oligosaccharides, especially their terminal sialic acid of GAT1 are necessary for the function of GAT1, their effects on the modification of N-glycans of GAT1 were also determined by the quantitative analysis of the sialic acid of GAT1. The results showed that 10 mmol /L 3-O-methyl-N-acetylglucosamine(3-O-Met-GlcNAc)reduced the GABA uptake activity of GAT1 to 53%(P< 0. 01)through inhibiting the sialylation of GAT1(66. 8%). And the GABA uptake activities were decreased to 54%, 63%, 63% and 67%(P< 0. 01), respectively, by the treatments of 10 mmol /L N-propionylglucosamine(GlcNProp), N-hexanoylglucosamine(GlcNHex), N-cycloproylgormylglucosamine(GlcNCyclo)and N-acetamidoacetylglucosamine(GlcNAc-acetamido)through inhibiting N-glycan trimming. These results indicate that the analogues of glucosamines have great potential in the development of inhibitors of GAT1 activity or sialic acid biosynthesis.
    • HTML全文
    • 参考文献(17)
  • [1] Hu J,Ouick MW.Substrate-mediated regulation of γ-aminobutyric acid transporter 1 in rat brain[J].Neuropharmacology,2008,54(2):309-318.
    [2] Chiu CS,Brickley SG,Jensen K,et al.GABA transporter deficiency causes tremor,ataxia,nervousness,and increased GABA-induced tonic conductance in cerebellum[J].J Neurosci,2005,25(12):3234-3245.
    [3] Bowery NG,Smart TG.GABA and glycine as neurotransmitters:a brief history[J].Br J Pharmacol,2006,147(S1):S109-S119.
    [4] Radian R,Kanner BI.Stoichiometry of sodium- and chloride-coupled γ-aminobutyric acid transport by synaptic plasma membrane vesicles isolated from rat brain[J].Biochemistry,1983,22:1236-1241.
    [5] Liu QR,Lopez-Corcuera B,Mandiyan S,et al.Molecular characterization of four pharmacologically distinct gamma-aminobutyric acid transporters in mouse brain[J].J Biol Chem,1993,268:2106-2112.
    [6] Guastella J,Nelson N,Nelson H,et al.Cloning and expression of a rat brain GABA transporter[J].Science,1990,249(4974):1303-1306.
    [7] Grossman TR,Nelson N.Effect of sodium lithium and proton concentrations on the electrophysiological properties of the four mouse GABA transporters expressed in Xenopus oocytes[J].Neurochem Int,2003,43(4/5):431-443.
    [8] Cope DW, Di Giovanni G, Fyson SJ, et al. Enhanced tonic GABAA inhibition in typical absence epilepsy[J].Nat Med,2009,15(12):1392-1398.
    [9] Liu GX,Liu S,Cai GQ,et al.Reduced aggression in mice lacking GABA transporter subtype 1[J].J Neurosci Res,2007,85(3):649-655.
    [10] Garcia-Alloza M,Tsang SW,Gil-Bea FJ,et al.Involvement of the GABAergic system in depressive symptoms of Alzheimer′s disease[J].Neurobiol Aging,2006,27(8):1110-1117.
    [11] Jasmin L,Wu MV,Ohara PT.GABA puts a stop to pain[J].Curr Drug Targets CNS Neurol Disord,2004,3(6):487-505.
    [12] Cai GQ,Salonikidis PS,Fei J,et al.The role of N-glycosylation in the stability,trafficking and GABA-uptake of GABA-transporter 1.Terminal N-glycans facilitate efficient GABA-uptake activity of the GABA transporter[J].FEBS J,2005,272(7):1625-1638.
    [13] Hu J,Fei J,Reutter W,et al.Involvement of sialic acid in the regulation of γ-aminobutyric acid uptake activity of γ-aminobutyric acid transporter 1[J].Glycobiology,2011,21(3):329-339.
    [14] Chou WK,Hinderlich S,Reutter W,et al.Sialic acid biosynthesis:stereochemistry and mechanism of the reaction catalyzed by the mammalian UDP-N-acetylglucosamine 2-epimerase[J].J Am Chem Soc,2003,125(9):2455-2461.
    [15] Grunholz HJ,Harms E,Opetz M,et al.Inhibition of in vitro biosynthesis of N-acetylneuraminic acid by N-acyl- and N-alkyl-2-amino-2-deoxyhexoses[J].Carbohydr Res,1981,96(2):259-270.
    [16] Reutter W,BauerC.Inhibitors of glycoprotein biosynthesis[J].Adv Enzyme Regul,1985,24(6):405-416.
    [17] Zeitler R,Giannis A,Danneschewski S,et al.Inhibition of N-acetylglucosamine kinase and N-acetylmannosamine kinase by 3-O-methyl-N-acetyl-D-glucosamine in vitro[J].Eur J Biochem,1992,204(3):1165-1168.
    • 相关文章
    • 施引文献
    • 资源附件(0)
计量
  • 文章访问数:  842
  • HTML全文浏览量:  0
  • PDF下载量:  1645
  • 被引次数: 0
出版历程
  • 刊出日期:  2016-04-24

目录

摘要
HTML全文
    参考文献(17)
    相关文章
    施引文献
    资源附件(0)

    /

    返回文章
    返回

    版权所有：《中国药科大学学报》编辑部苏ICP备05007142号

    地址：江苏省南京市鼓楼区童家巷24号（210009）电话: 025-83271566E-mail: xuebao@cpu.edu.cn

    本系统由 北京仁和汇智信息技术有限公司 开发  百度统计

    访问量：311502

    x 关闭 永久关闭