The L530R variation associated with recurrent kidney stones impairs the structure and function of TRPV5.
The L530R variation associated with recurrent kidney stones impairs the structure and function of TRPV5.
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DOI：
10.1016/j.bbrc.2017.08.102
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发表时间：
2017-10-21
影响因子：
3.1
通讯作者：
Peng JB
中科院分区：
文献类型：
--
作者：
Wang L;Holmes RP;Peng JB
TRPV 5是一种钙离子选择性通道，在肾脏钙离子重吸收中起关键作用。最近，TRPV 5的一种罕见的L530 R变异（rs757494578）被发现与创始人群中的复发性肾结石相关。然而，目前还不清楚这种变异在多大程度上改变了TRPV 5的结构和功能。为了评估TRPV 5变体的功能和表达，进行爪蟾卵母细胞中的Ca 2+摄取和蛋白质印迹分析。L530 R变异使TRPV 5在爪蟾卵母细胞中的Ca ~（2+）摄取活性消失。表达的变体蛋白质具有复杂糖基化的急剧减少。为了评估这种L530 R变体的结构效应，基于TRPV 6的晶体结构对TRPV 5进行建模，并进行分子动力学模拟。模拟结果表明，L530 R变异破坏了L530和L502之间的疏水相互作用，破坏了跨膜结构域5的二级结构。这种变化也改变了它与膜脂质分子的相互作用。与电中性的L530相比，带正电荷的R530残基使表面静电势向正移动。R530被吸引到带负电荷的磷酸基团，而不是膜脂质的疏水碳原子。这使R530所在的孔螺旋和Ca 2+选择性过滤器中的D542残基向膜表面移动。这些改变可能导致TRPV 5的错误折叠、通道向质膜的易位减少和/或通道的Ca 2+转运功能受损，并最终破坏TRPV 5介导的Ca 2+重吸收。
TRPV5 is a Ca2+-selective channel that plays a key role in the reabsorption of Ca2+ ions in the kidney. Recently, a rare L530R variation (rs757494578) of TRPV5 was found to be associated with recurrent kidney stones in a founder population. However, it was unclear to what extent this variation alters the structure and function of TRPV5. To evaluate the function and expression of the TRPV5 variant, Ca2+ uptake in Xenopus oocytes and western blot analysis were performed. The L530R variation abolished the Ca2+ uptake activity of TRPV5 in Xenopus oocytes. The variant protein was expressed with drastic reduction in complex glycosylation. To assess the structural effects of this L530R variation, TRPV5 was modeled based on the crystal structure of TRPV6 and molecular dynamics simulations were carried out. Simulation results showed that the L530R variation disrupts the hydrophobic interaction between L530 and L502, damaging the secondary structure of transmembrane domain 5. The variation also alters its interaction with membrane lipid molecules. Compared to the electroneutral L530, the positively charged R530 residue shifts the surface electrostatic potential towards positive. R530 is attracted to the negatively charged phosphate group rather than the hydrophobic carbon atoms of membrane lipids. This shifts the pore helix where R530 is located and the D542 residue in the Ca2+-selective filter towards the surface of the membrane. These alterations may lead to misfolding of TRPV5, reduction in translocation of the channel to the plasma membrane and/or impaired Ca2+ transport function of the channel, and ultimately disrupt TRPV5-mediated Ca2+ reabsorption.
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影响因子：
9.8
作者：
Akey JM;Eberle MA;Rieder MJ;Carlson CS;Shriver MD;Nickerson DA;Kruglyak L
通讯作者：
Kruglyak L
DOI：
10.1152/ajprenal.90771.2008
发表时间：
2009-05-01
影响因子：
4.2
作者：
Na, Tao;Zhang, Wei;Peng, Ji-Bin
通讯作者：
Peng, Ji-Bin
影响因子：
2.9
作者：
Wang, Lingyun;Murphy-Ullrich, Joanne E.;Song, Yuhua
通讯作者：
Song, Yuhua
上皮钙通道TRPV6的晶体结构。
影响因子：
64.8
作者：
Saotome K;Singh AK;Yelshanskaya MV;Sobolevsky AI
通讯作者：
Sobolevsky AI
DOI：
10.1007/978-3-642-54215-2_13
发表时间：
2014-01-01
期刊：
MAMMALIAN TRANSIENT RECEPTOR POTENTIAL (TRP) CATION CHANNELS, VOL I
影响因子：
--
作者：
Na, Tao;Peng, Ji-Bin
通讯作者：
Peng, Ji-Bin