NRFL-1, the C. elegans NHERF orthologue, interacts with amino acid transporter 6 (AAT-6) for age-dependent maintenance of AAT-6 on the membrane.
NRFL-1, the C. elegans NHERF orthologue, interacts with amino acid transporter 6 (AAT-6) for age-dependent maintenance of AAT-6 on the membrane.
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DOI：
10.1371/journal.pone.0043050
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Kanai Y
中科院分区：
文献类型：
--
作者：
Hagiwara K;Nagamori S;Umemura YM;Ohgaki R;Tanaka H;Murata D;Nakagomi S;Nomura KH;Kage-Nakadai E;Mitani S;Nomura K;Kanai Y
Na+/H+交换调节因子（Na+/H+ exchange regulatory factor，NHERF）家族在调节跨膜蛋白在质膜上的定位和滞留中起着关键作用。由于家族成员之间的高度同源性，潜在的功能补偿一直是一个问题，在整理出个人NHERF号码的功能。在这里，我们学习了C。elegans NRFL-1（C01F6.6）（nherf-like protein 1），唯一的C.线虫是NHERF家族的直向同源物，这使得蠕虫成为具有NHERF同源物的低遗传冗余的模型。整合C.的生物信息学知识。通过将线虫蛋白与酵母双杂交，我们鉴定出NRFL-1是C. elegans蛋白中AAT-6的相互作用子。氨基酸转运蛋白（AAT）家族。GST下拉试验，定位研究和免疫共沉淀的组合证实了结合，并表征了PDZ相互作用。AAT-6定位于腔膜，即使在NRFL-1的情况下，当蠕虫是长达4天的年龄。光漂白后的荧光恢复（FRAP）分析表明，NRFL-1将AAT-6固定在管腔膜处。相反，当nrfl-1缺陷蠕虫为6天或更大时，未观察到AAT-6的膜定位，而AAT-6在具有NRFL-1的蠕虫中紧密定位于膜。对C.通过对Elegans NHERF蛋白的研究，我们发现NRFL-1是AAT-6的PDZ相互作用物，负责AAT-6在肠腔膜上的固定和年龄依赖性维持。
The NHERF (Na+/H+ exchanger regulatory factor) family has been proposed to play a key role in regulating transmembrane protein localization and retention at the plasma membrane. Due to the high homology between the family members, potential functional compensations have been a concern in sorting out the function of individual NHERF numbers. Here, we studied C. elegans NRFL-1 (C01F6.6) (nherf-like protein 1), the sole C. elegans orthologue of the NHERF family, which makes worm a model with low genetic redundancy of NHERF homologues. Integrating bioinformatic knowledge of C. elegans proteins into yeast two-hybrid scheme, we identified NRFL-1 as an interactor of AAT-6, a member of the C. elegans AAT (amino acid transporter) family. A combination of GST pull-down assay, localization study, and co-immunoprecipitation confirmed the binding and characterized the PDZ interaction. AAT-6 localizes to the luminal membrane even in the absence of NRFL-1 when the worm is up to four-day old. A fluorescence recovery after photobleaching (FRAP) analysis suggested that NRFL-1 immobilizes AAT-6 at the luminal membrane. When the nrfl-1 deficient worm is six-day or older, in contrast, the membranous localization of AAT-6 is not observed, whereas AAT-6 tightly localizes to the membrane in worms with NRFL-1. Sorting out the in vivo functions of the C. elegans NHERF protein, we found that NRFL-1, a PDZ-interactor of AAT-6, is responsible for the immobilization and the age-dependent maintenance of AAT-6 on the intestinal luminal membrane.
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DOI：
10.1083/jcb.101.4.1532
发表时间：
1985-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Francis GR;Waterston RH
通讯作者：
Waterston RH
DOI：
10.1006/bbrc.2000.2260
发表时间：
2000-03-05
影响因子：
3.1
作者：
Gengyo-Ando, K;Mitani, S
通讯作者：
Mitani, S
影响因子：
21.3
作者：
Gloerich, Martijn;ten Klooster, Jean Paul;Bos, Johannes L.
通讯作者：
Bos, Johannes L.
影响因子：
4.8
作者：
Kitagawa, Hiroshi;Izumikawa, Tomomi;Sugahara, Kazuyuki
通讯作者：
Sugahara, Kazuyuki
影响因子：
5.5
作者：
Donowitz, M;Cha, BY;Li, XH
通讯作者：
Li, XH