School of Biological Sciences, The University of Hong Kong, Pokfulam, Hong Kong, China.
Plant Signal Behav. 2012 Aug;7(8):927-32. doi: 10.4161/psb.20769. Epub 2012 Jul 25.
To date, Arabidopsis purple acid phosphatase 2 (AtPAP2) is the only known plant protein that is dual-targeted to chloroplasts and mitochondria by a C-terminal targeting signal. Using in vitro organelle import and green fluorescence protein (GFP) localization assays, we showed that AtPAP2 is located on, but not imported across the outer membrane (OM) of chloroplasts and mitochondria and exposed its N-terminal enzymatic domain to the cytosol. It was also found that a short stretch of 30 amino acids (a.a.) at the C-terminal region (a.a. 615-644) that contains a stretch of 18 hydrophobic residues, a WYAK motif and 8 hydrophilic residues is sufficient for dual-targeting. Mutation of WYAK to WYAE had no effect on dual-targeting ability suggesting that the charge within this flanking region alone is not an important determinant for dual-targeting.
迄今为止,拟南芥紫色酸性磷酸酶 2(AtPAP2)是唯一已知的通过 C 末端靶向信号双重靶向定位于叶绿体和线粒体的植物蛋白。通过体外细胞器导入和绿色荧光蛋白(GFP)定位测定,我们表明 AtPAP2 位于叶绿体和线粒体的外膜(OM)上,但不穿过该膜,并且将其 N 端酶结构域暴露于细胞质中。还发现 C 末端区域(a.a.615-644)的 30 个氨基酸(a.a.)短片段包含一段 18 个疏水性残基、WYAK 基序和 8 个亲水性残基,足以进行双重靶向。将 WYAK 突变为 WYAE 对双重靶向能力没有影响,这表明侧翼区域内的电荷本身不是双重靶向的重要决定因素。