Trafficking of GFP-AQP5 chimeric proteins conferred with unphosphorylated amino acids at their PKA-target motif ((152)SRRTS) in MDCK-II cells.
Trafficking of GFP-AQP5 chimeric proteins conferred with unphosphorylated amino acids at their PKA-target motif ((152)SRRTS) in MDCK-II cells.
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DOI：
10.2152/jmi.56.55
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发表时间：
2009-02
期刊：
影响因子：
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通讯作者：
M. Karabasil;T. Hasegawa;A. Azlina;N. Purwanti;Javkhlan Purevjav;C. Yao;T. Akamatsu;K. Hosoi
中科院分区：
文献类型：
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作者：
M. Karabasil;T. Hasegawa;A. Azlina;N. Purwanti;Javkhlan Purevjav;C. Yao;T. Akamatsu;K. Hosoi
制备了3个在AQP 5（一种外分泌型水通道）的（152）SRRTS处具有突变的PKA靶基序的构建体，并将其融合到绿色荧光蛋白cDNA的C端，以检测阻断（152）SRRTS（AQP 5的共有PKA靶基序）处的磷酸化对Madin-Darby犬肾-II（MDCK-II）细胞中表达的嵌合蛋白的转运或运输的影响。H-89处理增加了野生型GFP-AQP 5向顶膜的易位。所有3种突变分子的易位比对照野生型GFP-AQP 5多1.5至2倍。秋水仙素而不是细胞松弛素B抑制野生型GFP-AQP 5的易位。目前的研究结果表明，这个共识序列的去磷酸化增加GFP-AQP 5的易位，微管，而不是微丝参与了这一事件。
Three constructs having mutated PKA-target motif at (152)SRRTS of AQP5, an exocrine type water channel, were prepared and fused to C-terminus of green fluorescence protein cDNA to examine the effects of blocking of phosphorylation at (152)SRRTS (a consensus PKA-target motif of AQP5) on translocation or trafficking of the chimeric proteins expressed in the Madin-Darby canine kidney-II (MDCK-II) cells. H-89 treatment increased translocation of wild-type GFP-AQP5 to the apical membrane. All 3 mutant molecules translocated 1.5 to 2 times more than the control wild-type GFP-AQP5. Colchicine but not cytochalasin B inhibited the translocation of wild-type GFP-AQP5. Present results suggest dephosphorylation of this consensus sequence increase GFP-AQP5 translocation, and that microtubules but not microfilaments are involved in this event.