权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Analysis of the vectorial transport of amino acid and drugs in epithelial cells.

上皮细胞中氨基酸和药物的载体运输分析。

基本信息

批准号：
12144201
负责人：
SUGIYAMA Yuichi
金额：
$ 32.77万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12144201/
关键词：
transcellular transport epithelial cells vectorial transport ABC transporter biliary excretion organic anion amino acid transporter 尿細管腎臓 OATP2 MRP2 極性細胞上皮輸送膜ソーティング蛋白間相互作用 MDCK細胞

项目摘要

We investigated multiplicity of the transporters involved in vectorial transport of amino acids and drugs in epithelial cells as well as the regulatory mechanism of their membrane trafficking. The region of BCRP necessary for its apical sorting was identified. Contribution of transporters to the drug-transport was investigated in kidney and choroid plexus (OAT1 and OAT3, and PEPT2 andOAT3, respectively). Functional characterization of MRP3 and BCRP was performed, and their involvement in drug-transport in the small intestine has been suggested. Uptake (OATP2) and efflux (MRP2) transporters were co-expressed in a polarized cell line (MDCK II) as an in vitro model of hepatobiliary transport of organic anions. In vitro. transcellular transport across P-gp-expressing monolayers can be used to predict the extent to which intestinal absorption and brain uptake is affected by Pigp. Furthermore, using a double transfectant (Oatp4/Mrp2), in vitro transcellular transport can be used to predict h … More epatobiliary clearance of organic anions. Double-transfectants expressing Ntcp and Bsep was established as model of hepatobiliary transport of bile acids.Seven amino acid transporters (Asc-2, AGT1, TAT1, LAT3, LAT4, CAT5 and B^0AT1) were newly cloned, and their functional characterization was performed. As luminal transporter for organic anions in the kidney, OATv1, URAT1 and OAT7 were cloned, and their functional characterization was performed. Mutations in B0AT1 and URAT1 cause Hartnup disorder and renal hypouricemia, respectively. Transmembrane domain of rBAT and 4F2hc is necessary for its interaction with their partner molecules. The "VVPP" sequence at C-terminus of BAT1 is essential for its membrane sorting. RACK1 was isolated by yeast-two-hybrid using the sequence as bait. RACK1 is a scaffold-protein contains multiple PDZ motifs, and we confirmed that it interacts with "VVPP" sequence of BAT1. The PDZ interacting motif at the C-terminus of URAT1 interacts with PDZK1. PDZ interactions may play an important role in clustering transporters. Less

我们研究了上皮细胞中参与氨基酸和药物载体转运的转运蛋白的多样性及其膜转运的调控机制。确定了其顶端分选所需的BCRP区域。在肾脏和脉络丛中研究转运蛋白对药物转运的贡献（分别为OAT 1和OAT 3，以及PEPT 2和OAT 3）。对MRP 3和BCRP进行了功能表征，并表明它们参与了小肠中的药物转运。在极化细胞系（MDCK II）中共表达摄取（OATP 2）和外排（MRP 2）转运蛋白，作为有机阴离子肝胆转运的体外模型。体外跨P-gp表达单层的跨细胞转运可用于预测肠吸收和脑摄取受Pigp影响的程度。此外，使用双转染子（Oatp 4/Mrp 2），体外跨细胞转运可用于预测细胞凋亡。 ...更多信息有机阴离子的上胆汁清除。建立了表达Ntcp和Bsep的双转染细胞作为胆汁酸肝胆转运的模型，并克隆了7个新的氨基酸转运蛋白（Asc-2，AGT 1，TAT 1，LAT 3，LAT 4，CAT 5和B^0AT1），并对其功能进行了研究。作为肾脏中有机阴离子的腔转运蛋白，OATv 1、URAT 1和OAT 7被克隆，并进行了功能表征。B 0AT 1和URAT 1的突变分别引起Hartnup病和肾性低尿酸血症。rBAT和4F 2 hc的跨膜结构域是rBAT和4F 2 hc与它们的伴侣分子相互作用所必需的。BAT 1的C端的“VVPP”序列是其膜分选所必需的。以该序列为诱饵，利用酵母双杂交技术分离RACK 1。RACK 1是一个含有多个PDZ基序的支架蛋白，我们证实它与BAT 1的“VVPP”序列相互作用。URAT 1 C末端的PDZ相互作用基序与PDZK 1相互作用。PDZ相互作用可能在聚集转运蛋白中起重要作用。少