Distinct subcellular localization of transiently expressed types 1 and 2 iodothyronine deiodinases as determined by immunofluorescence confocal microscopy.
Distinct subcellular localization of transiently expressed types 1 and 2 iodothyronine deiodinases as determined by immunofluorescence confocal microscopy.
复制标题
DOI：
10.1210/endo.141.11.7872
复制
发表时间：
2000-11
期刊：
影响因子：
4.8
通讯作者：
M. Baqui;B. Gereben;J. Harney;P. R. Larsen;Antonio C. Bianco
中科院分区：
文献类型：
--
作者：
M. Baqui;B. Gereben;J. Harney;P. R. Larsen;Antonio C. Bianco
我们比较了flag表位标记的1型和2型脱碘酶（D1和D2）在人胚胎肾（HEK-293）和小鼠神经母细胞瘤（NB2A）细胞中短暂表达的亚细胞定位。D2是一种完整的膜蛋白，基于对pH 11下提取的抗性，其NH2端位于内质网（ER）。使用抗flag和抗GRP78/BiP抗体的免疫荧光共聚焦显微镜显示，FLAG-D1信号位于细胞外周，与ER特异性标记物GRP78/BiP不共定位。另一方面，在与GRP78/BiP蛋白共定位的内质网中发现了FLAG-D2蛋白。这些差异分布模式表明D1和D2的亚细胞分选是由内在蛋白序列决定的，可以解释D2生成的T3进入细胞核的通道。
We compared the subcellular localization of FLAG-epitope tagged Types 1 and 2 deiodinases (D1 and D2) transiently expressed in human embryonic kidney (HEK-293) and mouse neuroblastoma (NB2A) cells. D2 is an integral membrane protein based on resistance to extraction at pH 11 with the NH2 terminus in the endoplasmic reticulum (ER). Immunofluorescence confocal microscopy using anti-FLAG and anti-GRP78/BiP antibodies showed the FLAG-D1 signal was found in the periphery of the cells and not co-localized with the ER specific marker GRP78/BiP. On the other hand, FLAG-D2 protein was found in the ER co-localized with the GRP78/BiP protein. These differential distribution patterns indicate subcellular sorting of D1 and D2 is determined by intrinsic protein sequence and can explain the ready access of D2-generated T3 to the nucleus.