Membrane-anchored prolyl hydroxylase with an export signal from the endoplasmic reticulum.
Membrane-anchored prolyl hydroxylase with an export signal from the endoplasmic reticulum.
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DOI：
10.1111/j.1365-313x.2004.02279.x
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发表时间：
2004-11
期刊：
影响因子：
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通讯作者：
K. Yuasa;K. Toyooka;H. Fukuda;K. Matsuoka
中科院分区：
文献类型：
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作者：
K. Yuasa;K. Toyooka;H. Fukuda;K. Matsuoka
根据表达序列标签信息，从烟草BY-2细胞中克隆了一个新的Pro-4-羟基酶(PH；EC 1.14.11.2)同源基因。与其他PHS一样，该烟草PH多肽含有两个保守的组氨酸残基，由286个氨基酸组成，计算相对分子质量为32 kDa。有趣的是，这种蛋白质及其在拟南芥和水稻中的同源物预测了它们N-末端的跨膜序列。该PH同源物在BY-2细胞中以组氨酸标记蛋白的形式表达，表达的蛋白具有PH活性。膜与高盐、尿素和蛋白水解酶的孵育和去污剂的作用表明，该蛋白是一种完整的膜蛋白，具有II型构型。它的膜锚定性质是植物特有的，因为在动物身上还没有发现完整的膜PH。膜分离研究和免疫细胞化学研究表明，该蛋白定位于内质网和高尔基体。对该蛋白与绿色荧光蛋白融合的分析表明，该蛋白胞质N末端的碱性氨基酸在其从内质网输出中起作用。
We cloned a novel prolyl 4-hydroxylase (PH; EC 1.14.11.2) homolog cDNA from tobacco (Nicotiana tabacum) BY-2 cells based on expression sequence tag information. Like other PHs, this tobacco PH polypeptide has two conserved histidine residues, and it comprises 286 amino acids with a calculated molecular mass of 32 kDa. Interestingly, this protein and homologs in Arabidopsis and rice have predicted transmembrane sequences in their N-terminal regions. This PH homolog was expressed in BY-2 cells as a His-tagged protein, and the expressed protein showed PH activity. Incubation of membranes with high salt, urea, and protease with or without detergents indicated that this protein is an integral membrane protein with a type II configuration. Its membrane-anchored nature is specific for plants because no integral membrane PH has been found in animals. A membrane fractionation study and immunocytochemical studies indicate that this protein localizes in both the endoplasmic reticulum (ER) and Golgi apparatus. Analysis of this protein fused to green fluorescent protein indicated that basic amino acids in the cytoplasmic, N-terminal region of the PH play a role in its export from the ER.