Mitochondrial 2′, 3′-cyclic nucleotide 3′-phosphodiesterase (CNP) interacts with mPTP modulators and functional complexes (I-V) coupled with release of apoptotic factors
Mitochondrial 2′, 3′-cyclic nucleotide 3′-phosphodiesterase (CNP) interacts with mPTP modulators and functional complexes (I-V) coupled with release of apoptotic factors
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DOI：
10.1016/j.neuint.2015.07.012
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发表时间：
2015-11-01
影响因子：
4.2
通讯作者：
Reiser, Georg
中科院分区：
文献类型：
--
作者：
Baburina, Yulia;Azarashvili, Tamara;Reiser, Georg
我们以前曾报道过2 '，3'-环核苷酸3 '-磷酸二酯酶（CNP）存在于大鼠脑和肝线粒体、外膜和线粒体质体中。发现CNP的底物2 '，3'-cAMP和2 '，3'-cNADP加速线粒体通透性转换孔（mPTP）的开放。在纯化的非突触线粒体中，观察到CNP与mPTP的主要调节剂（即VDAC、ANT和亲环素D）以及微管蛋白和考克斯IV共免疫沉淀。使用蓝色天然电泳，随后的蛋白质印迹，显示CNP与功能性内膜线粒体复合物I-V。在Ca 2+超载的线粒体中，CNP与复合物I-V的结合减少。环孢素A增加了CNP与复合物I和III的结合，支持这些复合物参与mPTP功能的想法。在Ca 2+超载的线粒体中（即当孔打开时），2 '，3'-cAMP增强CNP从复合物I、III、IV和V的解离。CNP与复合物I、III、IV和V的缔合在从脑、肝和心脏分离的线粒体中显示。线粒体中非选择性孔开放的刺激与CNP从线粒体释放相关，同时释放细胞色素c、AIF和Endo G。在Ca ~（2+）超载的线粒体中，2 '，3'-cAMP进一步促进AIF、Endo G和CNP的释放，但不影响细胞色素c的释放。这些结果为CNP作为mPTP复合物的可能调节因子之一参与线粒体呼吸和能量产生的调控提供了有力的证据。这揭示了CNP在髓鞘结构之外的新功能。(C)2015爱思唯尔有限公司版权所有。
We previously reported that 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP) is present in rat brain and liver mitochondria, in the outer membrane and mitoplasts. Substrates of CNP, 2',3'-cAMP and 2',3'-cNADP, were found to accelerate opening of mitochondrial permeability transition pore (mPTP). In purifled non-synaptic mitochondria, CNP was observed to co-immunoprecipitate with main modulators of mPTP, i.e. VDAC, ANT, and cyclophilin D, as well as with tubulin and COX IV. Using Blue Native Electrophoresis, with following Western blot, CNP was revealed to associate with functional inner membrane mitochondrial complexes I-V. In Ca2+-overloaded mitochondria, association of CNP with complexes I-V was decreased. Cyclosporine A increased the association of CNP with complexes I and III, supporting the idea of the involvement of these complexes in mPTP function. 2',3'-cAMP enhanced CNP dissociation from complexes I, III, IV and V in Ca2+-overloaded mitochondria (i.e. when pore is opened). Association of CNP with complexes I, III, IV, and V was shown in mitochondria isolated from brain, liver and heart. Stimulation of the opening of the non-selective pore in mitochondria correlated with CNP release from mitochondria in parallel with release of cytochrome c, AIF and Endo G. In Ca2+-overloaded mitochondria, 2',3'-cAMP further accelerated the release of AIF, Endo G and CNP, but did not alter cytochrome c release. These results provide strong evidence that CNP, one of the possible regulators of mPTP complex, might be involved in the control of respiration and energy production in mitochondria. This reveals a new function of CNP outside the myelin structure. (C) 2015 Elsevier Ltd. All rights reserved.