Identification of Bax-voltage-dependent anion channel 1 complexes in digitonin-solubilized cerebellar granule neurons.
Identification of Bax-voltage-dependent anion channel 1 complexes in digitonin-solubilized cerebellar granule neurons.
复制标题
在digiton-溶解小脑颗粒神经元中鉴定BAX-电压依赖性阴离子通道1复合物。
DOI：
10.1111/j.1471-4159.2011.07499.x
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发表时间：
2011-12
影响因子：
4.7
通讯作者：
Jekabsons MB
中科院分区：
文献类型：
--
作者：
Huckabee DB;Jekabsons MB
线粒体外膜Bax寡聚体对细胞色素c的释放至关重要，但线粒体蛋白在此过程中的作用仍不清楚。膜相关的Bax主要使用CHAPS作为增溶剂进行研究，因为它不会诱导构象伪影，尽管最近的证据表明它可能具有伪影效应。本研究的目的是研究毛地黄皂苷作为替代去污剂，以评估Bax寡聚状态，以及可能的相互作用与VDAC 1在小脑颗粒神经元。VDAC 1与Bax在来自健康和凋亡神经元的毛地黄皂苷提取物中共免疫沉淀。二维蓝色非变性SDS PAGE显示5个Bax和VDAC 1寡聚体具有相似的质量从120 - 500 kDa。Bax 1D 1免疫缺失提取物中两种VDAC 1寡聚体的水平与共沉淀的VDAC 1水平呈负相关，表明共沉淀的VDAC 1源自这些寡聚体。用6A7抗体的免疫耗竭适度地降低了来自凋亡但不健康神经元的Bax寡聚体的水平。第六个170 kDa的寡聚体只含有6A7 Bax和没有VDAC1后，细胞凋亡诱导。CHAPS不能溶解VDAC 1，并且另外没有产生明显的低聚物。我们得出结论，毛地黄皂苷是一种潜在的有用的去污剂，保留了可能被CHAPS破坏的PD-VDAC 1相互作用。
Mitochondrial outer membrane Bax oligomers are critical for cytochrome c release, but the role of resident mitochondrial proteins in this process remain unclear. Membrane-associated Bax has primarily been studied using CHAPS as the solubilizing agent, as it does not induce conformational artifacts, although recent evidence indicates it may have artifactual effects. The objective of this study was to investigate digitonin as an alternative detergent to assess Bax oligomeric state, and possible interaction with VDAC1 in cerebellar granule neurons. VDAC1 co-immunoprecipitated with Bax in digitonin extracts from healthy and apoptotic neurons. Two-dimensional blue native-SDS PAGE revealed five Bax and VDAC1 oligomers having similar masses from 120–500 kDa. The levels of two VDAC1 oligomers in Bax 1D1 immunodepleted extracts negatively correlated with levels of co-precipitated VDAC1, indicating the co-precipitated VDAC1 was derived from these oligomers. Immunodepletion with the 6A7 antibody modestly reduced the levels of Bax oligomers from apoptotic but not healthy neurons. A sixth 170 kDa oligomer containing exclusively 6A7 Bax and no VDAC1 was identified after apoptosis induction. CHAPS failed to solubilize VDAC1, and additionally yielded no distinct oligomers. We conclude that digitonin is a potentially useful detergent preserving Bax-VDAC1 interactions that may be disrupted with CHAPS.
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Bax缺失进一步命令小脑颗粒细胞中的细胞死亡途径，并提出了与caspase无关的细胞死亡途径。
影响因子：
7.8
作者：
Miller, T M;Moulder, K L;Knudson, C M;Creedon, D J;Deshmukh, M;Korsmeyer, S J;Johnson, E M Jr
通讯作者：
Johnson, E M Jr
DOI：
10.1073/pnas.94.8.3668
发表时间：
1997-04-15
影响因子：
11.1
作者：
Hsu, YT;Wolter, KG;Youle, RJ
通讯作者：
Youle, RJ
DOI：
10.1073/pnas.95.25.14681
发表时间：
1998-12-08
影响因子：
11.1
作者：
Narita, M;Shimizu, S;Tsujimoto, Y
通讯作者：
Tsujimoto, Y
影响因子：
4.8
作者：
Kim, Bong-Jo;Ryu, Seung-Wook;Song, Byoung-Joon
通讯作者：
Song, Byoung-Joon
影响因子：
5.3
作者：
Linseman, DA;Butts, BD;Heidenreich, KA
通讯作者：
Heidenreich, KA