Structural and biochemical consequences of disease-causing mutations in the ankyrin repeat domain of the human TRPV4 channel.
Structural and biochemical consequences of disease-causing mutations in the ankyrin repeat domain of the human TRPV4 channel.
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DOI：
10.1021/bi300279b
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发表时间：
2012-08-07
期刊：
影响因子：
2.9
通讯作者：
Gaudet R
中科院分区：
文献类型：
--
作者：
Inada H;Procko E;Sotomayor M;Gaudet R
TRPV4钙离子通透性通道在渗透调节、机械调节、细胞屏障形成和骨稳态等方面发挥着重要的生理作用。最近的研究报道，TRPV4的突变，包括其Ankyrin Repeat结构域(ARD)的一些突变，与人类遗传性疾病有关，包括神经疾病和骨骼发育不良，可能是因为该通道的结构活性增加。TRPV4的活性受钙调素和小分子如ATP与其胞质N端的ARD结合的调节。我们确定了人TRPV4-ARD的无ATP和结合形式的结构，并将它们与现有的TRPV-ARD结构进行了比较。第三重复间环区(指3环)是灵活的，并且可以充当调节通道活动的开关。对TRPV-ARD结构的比较也表明ARD结构与ATP结合能力之间存在进化联系。热稳定性分析和分子动力学模拟表明，ATP增加了能与ATP结合的TRPV-ARDS的稳定性。对与人类遗传性疾病相关的大量TRPV4-ARD突变进行的生化分析表明，一些突变损害了热稳定性，而另一些突变削弱了ATP结合能力，提示了这些疾病的分子机制。
The TRPV4 calcium-permeable cation channel plays important physiological roles in osmosensation, mechanosensation, cell barrier formation, and bone homeostasis. Recent studies reported that mutations in TRPV4, including some in its ankyrin repeat domain (ARD), are associated with human inherited diseases, including neuropathies and skeletal dysplasias, probably because of the increased constitutive activity of the channel. TRPV4 activity is regulated by the binding of calmodulin and small molecules such as ATP to the ARD at its cytoplasmic N-terminus. We determined structures of ATP-free and -bound forms of human TRPV4-ARD and compared them with available TRPV-ARD structures. The third inter-repeat loop region (Finger 3 loop) is flexible and may act as a switch to regulate channel activity. Comparisons of TRPV-ARD structures also suggest an evolutionary link between ARD structure and ATP binding ability. Thermal stability analyses and molecular dynamics simulations suggest that ATP increases stability in TRPV-ARDs that can bind ATP. Biochemical analyses of a large panel of TRPV4-ARD mutations associated with human inherited diseases showed that some impaired thermal stability while others weakened ATP binding ability, suggesting molecular mechanisms for the diseases.
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DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
4.8
作者：
Fecto, Faisal;Shi, Yong;Deng, Han-Xiang
通讯作者：
Deng, Han-Xiang
影响因子：
5.3
作者：
Alessandri-Haber, N;Dina, OA;Levine, JD
通讯作者：
Levine, JD
影响因子：
21.1
作者：
Clapham, DE;Montell, C;Julius, D
通讯作者：
Julius, D
影响因子：
4
作者：
Dai, J.;Kim, O-H;Ikegawa, S.
通讯作者：
Ikegawa, S.