Evaluation of sterol transport from the endoplasmic reticulum to mitochondria using mitochondrially targeted bacterial sterol acyltransferase in Saccharomyces cerevisiae.
Evaluation of sterol transport from the endoplasmic reticulum to mitochondria using mitochondrially targeted bacterial sterol acyltransferase in Saccharomyces cerevisiae.
复制标题
使用酿酒酵母中线粒体靶向细菌甾醇酰基转移酶评估甾醇从内质网到线粒体的转运。
DOI：
10.1080/09168451.2015.1058702
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
and R. Fukuda.
中科院分区：
文献类型：
--
作者：
S. Tian;A. Ohta;H. Horiuchi;and R. Fukuda.
为了阐明细胞器间固醇转运的机制，构建了一个评估内质网(ER)到线粒体的固醇转运的系统。将细菌甘油磷脂：胆固醇酰基转移酶与线粒体靶向序列和线粒体内膜蛋白Pet100的跨膜区以及增强型绿色荧光蛋白融合，在编码酰基-辅酶A：固醇酰基转移酶ARE1和ARE2缺失的酿酒酵母中表达。显微镜观察和亚细胞分离表明，该融合蛋白定位于线粒体中，命名为mito-SATA-EGFP。在表达Mito-SATA-EGFP的突变体中合成了甾醇酯。该系统将适用于通过检测线粒体中的甾醇酯化反应来评估内质网向酵母线粒体的甾醇运输。
To elucidate the mechanism of interorganelle sterol transport, a system to evaluate sterol transport from the endoplasmic reticulum (ER) to the mitochondria was constructed. A bacterial glycerophospholipid: cholesterol acyltransferase fused with a mitochondria-targeting sequence and a membrane-spanning domain of the mitochondrial inner membrane protein Pet100 and enhanced green fluorescent protein was expressed in aSaccharomyces cerevisiaemutant deleted forARE1andARE2encoding acyl-CoA:sterol acyltransferases. Microscopic observation and subcellular fractionation suggested that this fusion protein, which was named mito-SatA-EGFP, was localized in the mitochondria. Steryl esters were synthesized in the mutant expressing mito-SatA-EGFP. This system will be applicable for evaluations of sterol transport from the ER to the mitochondria in yeast by examining sterol esterification in the mitochondria.