Molecular interactions of the plasma membrane calcium ATPase 2 at pre- and post-synaptic sites in rat cerebellum.
Molecular interactions of the plasma membrane calcium ATPase 2 at pre- and post-synaptic sites in rat cerebellum.
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DOI：
10.1016/j.neuroscience.2009.04.059
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发表时间：
2009-08-18
期刊：
影响因子：
3.3
通讯作者：
Empson, R. M.
中科院分区：
文献类型：
--
作者：
Garside, M. L.;Turner, P. R.;Austen, B.;Strehler, E. E.;Beesley, P. W.;Empson, R. M.
质膜钙挤出机制，PMCA（质膜钙ATP酶）亚型2在脑中，特别是小脑中丰富表达。虽然已知PMCA 2与多种蛋白质相互作用以参与重要的信号传导事件，但其在脑突触组织中的分子相互作用尚不清楚。初步的蛋白质组学筛选和生化分离方法确定了PMCA 2和潜在的合作伙伴在突触前和突触后的网站从大鼠富集的脑组织。相互免疫沉淀和GST下拉方法证实PMCA 2通过其C-末端PDZ（PSD 95/Dlg/ZO-1）结合结构域与突触后蛋白PSD 95和NMDA谷氨酸受体亚基NR 1和NR 2a相互作用。由于PSD 95是NMDA受体的一个众所周知的伴侣，这就提出了一个令人兴奋的可能性，即所有三种相互作用都发生在同一个突触后信号复合物中。在突触前，其中PMCA 2存在于突触前网络中，相互免疫沉淀和GST下拉方法确定了突触前膜蛋白syntaxin-1A，SNARE复合物的成员，作为PMCA 2的潜在伴侣。PSD 95-PMCA 2和syntaxin-1A-PMCA 2的相互作用也被检测到的分子和颗粒细胞层的大鼠小脑矢状切片的免疫组织化学。这些特定的分子相互作用在小脑突触可能允许PMCA 2密切控制局部钙动力学的一部分，前和突触后信号复合物。
The plasma membrane calcium extrusion mechanism, PMCA (plasma membrane calcium ATPase) isoform 2 is richly expressed in the brain and particularly the cerebellum. Whilst PMCA2 is known to interact with a variety of proteins to participate in important signalling events, its molecular interactions in brain synapse tissue are not well understood. An initial proteomics screen and a biochemical fractionation approach identified PMCA2 and potential partners at both pre- and post-synaptic sites in synapse-enriched brain tissue from rat. Reciprocal immunoprecipitation and GST pull down approaches confirmed that PMCA2 interacts with the post-synaptic proteins PSD95 and the NMDA glutamate receptor subunits NR1 and NR2a, via its C-terminal PDZ (PSD95/Dlg/ZO-1) binding domain. Since PSD95 is a well known partner for the NMDA receptor this raises the exciting possibility that all three interactions occur within the same post-synaptic signalling complex. At the pre-synapse, where PMCA2 was present in the pre-synapse web, reciprocal immunoprecipitation and GST pull down approaches identified the pre-synaptic membrane protein syntaxin-1A, a member of the SNARE complex, as a potential partner for PMCA2. Both PSD95-PMCA2 and syntaxin-1A-PMCA2 interactions were also detected in the molecular and granule cell layers of rat cerebellar sagittal slices by immunohistochemistry. These specific molecular interactions at cerebellar synapses may allow PMCA2 to closely control local calcium dynamics as part of pre-and post-synaptic signalling complexes.
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DOI：
10.1073/pnas.0811025106
发表时间：
2008-12-30
影响因子：
11.1
作者：
Elias, G. M.;Elias, L. A. B.;Nicoll, R. A.
通讯作者：
Nicoll, R. A.
影响因子：
3.5
作者：
Buckby, LE;Jensen, TP;Empson, RM
通讯作者：
Empson, RM
影响因子：
5.3
作者：
Kim, MH;Korogod, N;Lee, SH
通讯作者：
Lee, SH
影响因子：
5.3
作者：
Empson, Ruth M.;Garside, Molly L.;Knopfel, Thomas
通讯作者：
Knopfel, Thomas
影响因子：
4.8
作者：
DeMarco, SJ;Chicka, MC;Strehler, EE
通讯作者：
Strehler, EE