A cytosolic phospholipase A2-like protein in the filamentous fungus Aspergillus oryzae localizes to the intramembrane space of the mitochondria.
A cytosolic phospholipase A2-like protein in the filamentous fungus Aspergillus oryzae localizes to the intramembrane space of the mitochondria.
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DOI：
10.1111/j.1574-6968.2009.01818.x
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发表时间：
2009-12
影响因子：
2.1
通讯作者：
Kohei Takaya;Yujiro Higuchi;K. Kitamoto;M. Arioka
中科院分区：
文献类型：
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作者：
Kohei Takaya;Yujiro Higuchi;K. Kitamoto;M. Arioka
在哺乳动物细胞中，胞质磷脂酶A(2) （cPLA(2)）在细胞刺激下通过释放花生四烯酸表现出多种活性。在这项研究中，我们对丝状真菌米曲霉中可能存在的cPLA(2)样蛋白AoPlaA进行了表征。当AoPlaA-EGFP在m.o oryzae中表达时，它定位于线粒体标记染料染色的管状结构上。生化分离实验表明，线粒体富集部位存在AoPlaA。通过n端氨基酸序列分析，证实了AoPlaA中存在一个n端可切割的线粒体靶向信号，并发现由AoPlaA n端65或50个氨基酸组成的嵌合蛋白与定位于线粒体的增强型绿色荧光蛋白（EGFP）融合。在酿酒酵母中表达的AoPlaA的亚线粒体分离表明，AoPlaA定位于线粒体的膜内空间。综上所述，本文提出的结果证明了丝状真菌中cPLA(2)样蛋白的一种新的亚线粒体定位。
In mammalian cells, cytosolic phospholipase A(2) (cPLA(2)) displays a variety of activities through the release of arachidonic acid in response to cellular stimuli. In this study, we characterize the putative cPLA(2)-like protein, AoPlaA, in the filamentous fungus Aspergillus oryzae. When AoPlaA-EGFP was expressed in A. oryzae, it localized to the tubular structures that was costained by the marker dye for the mitochondria. A biochemical fractionation experiment showed that AoPlaA was present in the mitochondria-enriched fraction. The presence of an N-terminal cleavable mitochondrial targeting signal in AoPlaA was demonstrated by N-terminal amino acid sequence analysis, and by showing that chimeric proteins consisting of N-terminal 65 or 50 amino acids of AoPlaA fused to enhanced green fluorescent protein (EGFP) localized to the mitochondria. Submitochondrial fractionation of AoPlaA expressed in Saccharomyces cerevisiae demonstrated that AoPlaA localizes to the intramembrane space of the mitochondria. Taken together, the results presented here demonstrate a novel, submitochondrial localization of the cPLA(2)-like protein in the filamentous fungi.