Biochemical Characterization of Cone Cyclic Nucleotide-Gated (CNG) Channel Using the Infrared Fluorescence Detection System
Biochemical Characterization of Cone Cyclic Nucleotide-Gated (CNG) Channel Using the Infrared Fluorescence Detection System
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DOI：
10.1007/978-1-4614-0631-0_98
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发表时间：
2012-01-01
期刊：
影响因子：
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通讯作者：
Matsumoto, Hiroyuki
中科院分区：
文献类型：
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作者：
Ding, Xi-Qin;Matveev, Alexander;Matsumoto, Hiroyuki
视锥视觉是由光感受器环核苷门控通道介导的，它对中心视觉、色觉和视觉灵敏度至关重要。锥形CNG通道由两个构造相关的亚基类型CNGA3和CNGB3组成。自然发生的锥体CNG通道突变与多种锥体疾病有关，包括色盲、进行性锥体营养不良和一些黄斑病变。然而，我们对锥形CNG通道结构的认识相当有限。这在一定程度上是由于研究以杆状细胞为主的哺乳动物视网膜中的视锥细胞所面临的挑战。我们已经证明了锥体CNG通道的鲁棒性表达和杆状CNG通道的缺失，并表明Nrl - / -小鼠系是研究锥体CNG通道的有价值的模型。本研究采用红外荧光Western检测结合化学交联和蓝色原生page对锥形CNG通道的复杂结构进行了研究。我们的研究结果表明，天然锥体CNG通道是一种异四聚体复合物，其化学计量可能为3个CNGA3和1个CNGB3。
Cone vision mediated by photoreceptor cyclic nucleotide-gated (CNG) channel is essential for central and color vision and visual acuity. Cone CNG channel is composed of two structurally related subunit types, CNGA3 and CNGB3. Naturally occurring mutations in cone CNG channel are associated with a variety of cone diseases including achromatopsia, progressive cone dystrophy, and some maculopathies. Nevertheless, our understanding of the structure of cone CNG channel is quite limited. This is, in part, due to the challenge of studying cones in a rod-dominant mammalian retina. We have demonstrated a robust expression of cone CNG channel and a lack of rod CNG channel in the cone-dominant Nrl−/− retina and shown that the Nrl−/− mouse line is a valuable model to study cone CNG channel. This work examined the complex structure of cone CNG channel using infrared fluorescence Western detection combined with chemical cross-linking and blue native-PAGE. Our results suggest that the native cone CNG channel is a heterotetrameric complex likely at a stoichiometry of three CNGA3 and one CNGB3.