Center for Integrated Protein Science CIPS-M, Department Pharmazie, Ludwig-Maximilians-Universität München, D-81377 München, Germany.
J Biol Chem. 2012 Oct 19;287(43):36312-21. doi: 10.1074/jbc.M112.392811. Epub 2012 Aug 30.
Cav1.4 L-type Ca(2+) channels are crucial for synaptic transmission in retinal photoreceptors and bipolar neurons. Recent studies suggest that the activity of this channel is regulated by the Ca(2+)-binding protein 4 (CaBP4). In the present study, we explored this issue by examining functional effects of CaBP4 on heterologously expressed Cav1.4. We show that CaBP4 dramatically increases Cav1.4 channel availability. This effect crucially depends on the presence of the C-terminal ICDI (inhibitor of Ca(2+)-dependent inactivation) domain of Cav1.4 and is absent in a Cav1.4 mutant lacking the ICDI. Using FRET experiments, we demonstrate that CaBP4 interacts with the IQ motif of Cav1.4 and that it interferes with the binding of the ICDI domain. Based on these findings, we suggest that CaBP4 increases Cav1.4 channel availability by relieving the inhibitory effects of the ICDI domain on voltage-dependent Cav1.4 channel gating. We also functionally characterized two CaBP4 mutants that are associated with a congenital variant of human night blindness and other closely related nonstationary retinal diseases. Although both mutants interact with Cav1.4 channels, the functional effects of CaBP4 mutants are only partially preserved, leading to a reduction of Cav1.4 channel availability and loss of function. In conclusion, our study sheds new light on the functional interaction between CaBP4 and Cav1.4. Moreover, it provides insights into the mechanism by which CaBP4 mutants lead to loss of Cav1.4 function and to retinal disease.
Cav1.4 L 型钙 (Ca2+) 通道对于视网膜光感受器和双极神经元中的突触传递至关重要。最近的研究表明,该通道的活性受 Ca2+结合蛋白 4(CaBP4)的调节。在本研究中,我们通过检查 CaBP4 对异源表达的 Cav1.4 的功能影响来探讨这个问题。我们表明 CaBP4 显著增加了 Cav1.4 通道的可用性。这种作用主要取决于 Cav1.4 的 C 端 ICDI(Ca2+依赖性失活抑制剂)结构域的存在,并且在缺乏 ICDI 的 Cav1.4 突变体中不存在。通过 FRET 实验,我们证明 CaBP4 与 Cav1.4 的 IQ 基序相互作用,并且它干扰 ICDI 结构域的结合。基于这些发现,我们提出 CaBP4 通过解除 ICDI 结构域对电压依赖性 Cav1.4 通道门控的抑制作用来增加 Cav1.4 通道的可用性。我们还对与人类先天性夜盲症和其他密切相关的非静止性视网膜疾病相关的两种 CaBP4 突变体进行了功能表征。尽管这两种突变体都与 Cav1.4 通道相互作用,但 CaBP4 突变体的功能影响仅部分保留,导致 Cav1.4 通道的可用性降低和功能丧失。总之,我们的研究揭示了 CaBP4 与 Cav1.4 之间的功能相互作用。此外,它为 CaBP4 突变体导致 Cav1.4 功能丧失和视网膜疾病的机制提供了深入了解。