Extracellular Interaction of the Voltage-dependent Ca2+ Channel α2δ and α1 Subunits*

Extracellular Interaction of the Voltage-dependent Ca2+ Channel α2δ and α1 Subunits*
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电压依赖性 Ca2+ 通道 α2δ 和 α1 亚基的细胞外相互作用*

DOI:
--
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发表时间:
1997
影响因子:
4.8
通讯作者:
K. Campbell
K. Campbell
中科院分区:
生物学2区
文献类型:
--
作者:
C. Gurnett;R. Felix;K. Campbell

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研究了电压依赖性Ca ~(2+)通道α2δ亚基胞外区在与α 1C亚基组装中的作用。瞬时转染的tsA 201细胞适当地加工α2δ亚基,因为α2和δ亚基之间的二硫键和切割位点显示与天然通道蛋白相似。免疫共沉淀实验表明,在缺乏δ亚基的情况下,α2亚基不与α1亚基组装。此外,δ中的跨膜和胞质序列可以与不相关蛋白质的序列交换,而对α2δ和α1蛋白质之间的结合没有任何影响。α2δ亚基的细胞外结构域也被证明是增加[3 H] PN 200 -110(异丙基-4-(2,1,3-苯并恶二唑-4-基)-1,4-二氢-2,6-二甲基-5-([3 H]甲氧羰基)-吡啶-3-羧酸酯)与α1C亚基结合亲和力的原因。对α1亚基上相应相互作用位点的研究表明,尽管含有天然α 1 S亚基重复序列III的胰蛋白酶肽在麦胚凝集素层析过程中仍与α2δ亚基结合,但重复序列III本身不足以与α2δ亚基组装。我们的研究结果表明,α2δ亚基可能与α1亚基的一个以上的细胞外环相互作用。
The role of the extracellular domain of the voltage-dependent Ca2+ channel α2δ subunit in assembly with the α1Csubunit was investigated. Transiently transfected tsA201 cells processed the α2δ subunit properly as disulfide linkages and cleavage sites between the α2 and δ subunits were shown to be similar to native channel protein. Coimmunoprecipitation experiments demonstrated that in the absence of δ subunits, α2 subunits do not assemble with α1 subunits. Furthermore, the transmembrane and cytoplasmic sequences in δ can be exchanged with those of an unrelated protein without any effect on the association between the α2δ and α1 proteins. Extracellular domains of the α2δ subunit are also shown to be responsible for increasing the binding affinity of [3H]PN200-110 (isopropyl-4-(2,1,3-benzoxadiazol-4-yl)-1,4-dihydro-2,6-dimethyl-5-([3H]methoxycarbonyl)-pyridine-3-carboxylate) for the α1C subunit. Investigation of the corresponding interaction site on the α1 subunit revealed that although tryptic peptides containing repeat III of native α1S subunit remain in association with the α2δ subunit during wheat germ agglutinin chromatography, repeat III by itself is not sufficient for assembly with the α2δ subunit. Our results suggest that the α2δ subunit likely interacts with more than one extracellular loop of the α1 subunit.
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DOI: --
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DOI: --
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期刊: The Journal of biological chemistry
影响因子: --
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影响因子: 20.1
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通讯作者: GeorgeJr,AL