Measuring Ca2+-Dependent Modulation of Voltage-Gated Ca2+ Channels in HEK-293T Cells.
Measuring Ca2+-Dependent Modulation of Voltage-Gated Ca2+ Channels in HEK-293T Cells.
复制标题
测量 HEK-293T 细胞中电压门控 Ca2 通道的 Ca2 依赖性调制。
DOI：
10.1101/pdb.prot087213
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发表时间：
2016
影响因子：
--
通讯作者：
Lee,Amy
中科院分区：
文献类型：
--
作者：
Thomas,JessicaR;Lee,Amy
电压门控钙通道调节多种生物过程，如肌肉收缩、基因表达和神经递质释放。钙通道受到多种形式的调节，包括那些涉及渗透到毛孔中的钙离子的调节。高电压激活的钙通道经历钙依赖失活(CDI)和易化(CDF)，可调节心律和突触可塑性等过程。Cav1(L型)和Cav2(P/Q-、N-和R型)通道的CDI和CDF略有不同。SV40大T抗原(HEK-293T)转化的人胚胎肾细胞有利于研究特定类型钙通道的CDI和CDF。HEK-293T细胞不表达内源性钙通道，但瞬时转染法可高水平表达外源性钙通道。本方案描述了如何表征和分析HEK-293T细胞中重组钙通道的钙依赖调控。
Voltage-gated Ca 2+(Ca v) channels regulate a variety of biological processes, such as muscle contraction, gene expression, and neurotransmitter release. Ca v channels are subject to diverse forms of regulation, including those involving the Ca 2+ ions that permeate the pore. High voltage-activated Ca v channels undergo Ca 2+-dependent inactivation (CDI) and facilitation (CDF), which can regulate processes such as cardiac rhythm and synaptic plasticity. CDI and CDF differ slightly between Ca v 1 (L-type) and Ca v 2 (P/Q-, N-, and R-type) channels. Human embryonic kidney cells transformed with SV40 large T-antigen (HEK-293T) are advantageous for studying CDI and CDF of a particular type of Ca v channel. HEK-293T cells do not express endogenous Ca v channels, but Ca v channels can be expressed exogenously at high levels in these cells by transient transfection. This protocol describes how to characterize and analyze Ca 2+-dependent modulation of recombinant Ca v channels in HEK-293T cells.
膜、分子、毒素和细胞
影响因子：
0.9
作者：
R. Juliano
通讯作者：
R. Juliano