Acyl-chain remodeling of dioctanoyl-phosphatidylcholine in Saccharomyces cerevisiae mutant defective in de novo and salvage phosphatidylcholine synthesis
Acyl-chain remodeling of dioctanoyl-phosphatidylcholine in Saccharomyces cerevisiae mutant defective in de novo and salvage phosphatidylcholine synthesis
复制标题
酿酒酵母突变体中二辛酰磷脂酰胆碱的酰基链重塑缺陷从头和挽救磷脂酰胆碱合成
DOI：
10.1016/j.bbrc.2014.01.136
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发表时间：
2014
影响因子：
3.1
通讯作者：
Akinori Ohta
中科院分区：
文献类型：
--
作者：
Hideyuki Kishino;Hiroki Eguchi;Keiko Takagi;Hiroyuki Horiuchi;Ryouichi Fukuda;Akinori Ohta
利用编码磷脂酰乙醇胺甲基转移酶和磷脂甲基转移酶的双缺失突变体pem1和pem2中的gal1启动子替换编码CTP：磷脂酰胆碱转移酶的pct1启动子，构建了内源性磷脂酰胆碱合成可控的酵母菌株。该突变体不能在含葡萄糖的培养基中生长，但添加二辛烷酰磷脂酰胆碱（diC8PC）支持其生长。利用电喷雾串联质谱法对该菌株13c标记的diC8PC（（甲基- 13c）3-diC8PC）进行代谢分析，发现该菌株在sn-1和sn-2位置均转化为含有16或18个酰基残基的PC。此外，（甲基- 13c）3-diC8PC的两个酰基残基在棕榈酰辅酶a存在的酵母细胞提取物的体外反应中被16:1酰基链取代。这些结果表明含有短酰基残基的PC在酵母中被改造成具有生理长度的酰基链。
A yeast strain, in which endogenous phosphatidylcholine (PC) synthesis is controllable, was constructed by the replacement of the promoter ofPCT1, encoding CTP:phosphocholine cytidylyltransferase, withGAL1promoter in a double deletion mutant ofPEM1andPEM2, encoding phosphatidylethanolamine methyltransferase and phospholipid methyltransferase, respectively. This mutant did not grow in the glucose-containing medium, but the addition of dioctanoyl-phosphatidylcholine (diC8PC) supported its growth. Analyses of the metabolism of13C-labeled diC8PC ((methyl-13C)3-diC8PC) in this strain using electrospray ionization tandem mass spectrometry revealed that it was converted to PC species containing acyl residues of 16 or 18 carbons at bothsn-1 andsn-2 positions. In addition, both acyl residues of (methyl-13C)3-diC8PC were replaced with 16:1 acyl chains in thein vitroreaction using the yeast cell extract in the presence of palmitoleoyl-CoA. These results indicate that PC containing short acyl residues was remodeled to those with acyl chains of physiological length in yeast.