Role of ARF6 in internalization of metal-binding proteins, metallothionein and transferrin, and cadmium-metallothionein toxicity in kidney proximal tubule cells.
Role of ARF6 in internalization of metal-binding proteins, metallothionein and transferrin, and cadmium-metallothionein toxicity in kidney proximal tubule cells.
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ARF6 在金属结合蛋白、金属硫蛋白和转铁蛋白的内化以及肾近曲小管细胞中镉-金属硫蛋白毒性中的作用
DOI：
10.1016/j.taap.2008.02.008
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发表时间：
2008
影响因子：
3.8
通讯作者：
Thévenod
中科院分区：
文献类型：
--
作者：
Abouhamed;Thévenod
过滤后的金属蛋白复合物，如镉-金属硫蛋白-1 （CdMT-1）或转铁蛋白（Tf），在肾近端小管（PT）细胞中部分通过巨噬蛋白/立方蛋白被顶端内吞，CdMT-1内化导致细胞凋亡。小GTPase ARF （adp -核糖基化因子）蛋白调节内吞作用和囊泡运输。我们研究了ARF6在PT细胞摄取MT-1/CdMT-1和Tf后，参与PT细胞中配体内化和内吞运输的作用。来自大鼠PT S1片段的WKPT-0293 cl2细胞转染了血凝素标记的野生型（ARF6- wt）或显性阴性（ARF6- t27n）形式的ARF6。免疫荧光检测显示，内源性ARF6与质膜（PM）相关，并与Rab5a和Rab11共定位，参与早期和循环内体运输。免疫荧光染色显示ARF6显性阴性（ARF6- dn）细胞表面标记减少。ARF6-DN细胞内Alexa Fluor 546结合的MT-1摄取减少，CdMT-1 （14.8 μM / 24 h）毒性从ARF6-WT细胞的27.3±3.9%显著降低到ARF6-DN细胞的11.1±4.0% （n=6， P<0.02）。此外，在ARF-DN细胞中观察到Alexa Fluor 546共轭Tf摄取减少（75.0±4.6%，ARF6-WT细胞为3.9±3.9%,n=3， P<0.01）和/或保持在PM附近（89.3±5）。ARF6-WT细胞为45.2±14.3%,n=3， P<0.05)。综上所述，这些数据支持了ARF6在受体介导的PT细胞内吞作用、MT-1/Tf向内体/溶酶体转运以及CdMT-1毒性中的作用。
Filtered metal–protein complexes, such as cadmium-metallothionein-1 (CdMT-1) or transferrin (Tf) are apically endocytosed partly via megalin/cubilin by kidney proximal tubule (PT) cells where CdMT-1 internalization causes apoptosis. Small GTPase ARF (ADP-ribosylation factor) proteins regulate endocytosis and vesicular trafficking. We investigated roles of ARF6, which has been shown to be involved in internalization of ligands and endocytic trafficking in PT cells, following MT-1/CdMT-1 and Tf uptake by PT cells. WKPT-0293 Cl.2 cells derived from rat PT S1 segment were transfected with hemagglutinin-tagged wild-type (ARF6-WT) or dominant negative (ARF6-T27N) forms of ARF6. Using immunofluorescence, endogenous ARF6 was associated with the plasma membrane (PM) as well as juxtanuclear and co-localized with Rab5a and Rab11 involved in early and recycling endosomal trafficking. Immunofluorescence staining of megalin showed reduced surface labelling in ARF6 dominant negative (ARF6-DN) cells. Intracellular Alexa Fluor 546-conjugated MT-1 uptake was reduced in ARF6-DN cells and CdMT-1 (14.8 μM for 24 h) toxicity was significantly attenuated from 27.3±3.9% in ARF6-WT to 11.1±4.0% in ARF6-DN cells (n=6, P<0.02). Moreover, reduced Alexa Fluor 546-conjugated Tf uptake was observed in ARF-DN cells (75.0±4.6% versus 3.9±3.9% of ARF6-WT cells, n=3, P<0.01) and/or remained near the PM (89.3±5. 6% versus 45.2±14.3% of ARF6-WT cells, n=3, P<0.05). In conclusion, the data support roles for ARF6 in receptor-mediated endocytosis and trafficking of MT-1/Tf to endosomes/lysosomes and CdMT-1 toxicity of PT cells.
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DOI：
10.1152/ajprenal.00359.2005
发表时间：
2006-06-01
影响因子：
4.2
作者：
Abouhamed, M;Gburek, J;Smith, CP
通讯作者：
Smith, CP
DOI：
10.1111/j.1365-2362.1979.tb01662.x
发表时间：
1979-01-01
影响因子：
5.5
作者：
BERNARD, A;BUCHET, JP;LAUWERYS, R
通讯作者：
LAUWERYS, R
影响因子：
3.9
作者：
Donaldson, JG;Honda, A
通讯作者：
Honda, A
DOI：
10.1124/jpet.106.102574
发表时间：
2006-08
影响因子：
3.5
作者：
N. Wolff;Marouan Abouhamed;P. Verroust;F. Thévenod
通讯作者：
N. Wolff;Marouan Abouhamed;P. Verroust;F. Thévenod
影响因子：
19.6
作者：
P. Verroust;H. Birn;R. Nielsen;R. Kozyraki;E. Christensen
通讯作者：
P. Verroust;H. Birn;R. Nielsen;R. Kozyraki;E. Christensen