80K-H as a new Ca2+ sensor regulating the activity of the epithelial Ca2+ channel transient receptor potential cation channel V5 (TRPV5)

被引:56
作者
Gkika, D
Mahieu, F
Nilius, B
Hoenderop, JGJ
Bindels, RJM
机构
[1] Univ Nijmegen, Med Ctr, Nijmegen Ctr Mol Life Sci, Dept Physiol, NL-6500 HB Nijmegen, Netherlands
[2] Katholieke Univ Leuven, Dept Physiol, B-3000 Louvain, Belgium
关键词
D O I
10.1074/jbc.M403801200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The epithelial Ca2+ channel transient receptor potential cation channel V5 (TRPV5) constitutes the apical Ca2+ entry pathway in the process of active Ca2+ reabsorption. Ca2+ influx through TRPV5 is tightly controlled by modulators of Ca2+ homeostasis, including 1,25-dihydroxyvitamin D-3 and dietary Ca2+. However, little is known about intracellular proteins that interact with TRPV5 and directly regulate the activation of this channel. By the use of cDNA microarrays, the present study identified 80K-H as the first protein involved in the Ca2+-dependent control of the epithelial Ca2+ channel TRPV5. 80K-H was initially identified as a protein kinase C substrate, but its biological function remains to be established. We demonstrated a specific interaction between 80K-H and TRPV5, co-localization of both proteins in the kidney, and similar transcriptional regulation by 1,25-dihydroxyvitamin D3 and dietary Ca2+. Furthermore, 80K-H directly bound Ca2+, and inactivation of its two EF-hand structures totally abolished Ca2+ binding. Electrophysiological studies using 80K-H mutants showed that three domains of 80K-H (the two EF-hand structures, the highly acidic glutamic stretch, and the His-Asp-Glu-Leu sequence) are critical determinants for TRPV5 activity. Importantly, inactivation of the EF-hand pair reduced the TRPV5-mediated Ca2+ current and increased the TRPV5 sensitivity to intracellular Ca2+, accelerating the feedback inhibition of the channel. None of the 80K-H mutants altered the TRPV5 plasma membrane localization nor the association of 80K-H with TRPV5, suggesting that 80K-H has a direct effect on TRPV5 activity. In conclusion, we report a novel function for 80K-H as a Ca2+ sensor controlling TRPV5 channel activity.
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收藏
页码:26351 / 26357
页数:7
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