Identification and characterization of major lipid particle proteins of the yeast Saccharomyces cerevisiae
Identification and characterization of major lipid particle proteins of the yeast Saccharomyces cerevisiae
复制标题
DOI：
10.1128/jb.181.20.6441-6448.1999
复制
发表时间：
1999-10-01
影响因子：
3.2
通讯作者：
Daum, G
中科院分区：
文献类型：
--
作者：
Athenstaedt, K;Zweytick, D;Daum, G
从酿酒酵母（Saccharomycescerevisiae）中分离出高纯度的脂质颗粒，经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离其蛋白质，质谱分析鉴定了主要的脂质颗粒蛋白质，并推导了相应的开放阅读框架（ORF）。在silicio分析表明，所有的脂质颗粒蛋白含有几个疏水结构域，但没有或只有很少（假设）跨膜区。迄今为止，通过功能鉴定的所有脂质颗粒蛋白，如Erg1p、Erg6p和Erg7p（麦角固醇生物合成）和Faa1p、Faa4p和Fat1p（脂肪酸代谢），都参与脂质代谢。基于序列同源性，另一组三种脂质颗粒蛋白可能参与脂质降解。为了研究未知功能的脂质颗粒蛋白是否也参与脂质合成，构建了相应ORF缺失的突变体，并进行了系统的脂质分析。YDL 193 w的缺失导致致死表型，其不能通过补充麦角固醇或脂肪酸来抑制。其他缺失突变体在标准条件下是活的，YBR 177c、YMR 313c和YKL 140w缺失的菌株表现出不同于野生型菌株的磷脂和/或中性脂质模式，因此可能是参与酵母脂质代谢的进一步候选ORF。
Lipid particles of the yeast Saccharomyces cerevisiae were isolated at high purity, and their proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Major lipid particle proteins were identified by mass spectrometric analysis, and the corresponding open reading frames (ORFs) were deduced. In silicio analysis revealed that all lipid particle proteins contain several hydrophobic domains but none or only few (hypothetical) transmembrane spanning regions. All lipid particle proteins identified by function so far, such as Erg1p, Erg6p, and Erg7p (ergosterol biosynthesis) and Faa1p, Faa4p, and Fat1p (fatty acid metabolism), are involved in lipid metabolism. Based on sequence homology, another group of three lipid particle proteins may be involved in lipid degradation. To examine whether lipid particle proteins of unknown function are also involved in lipid synthesis, mutants with deletions of the respective ORFs were constructed and subjected to systematic lipid analysis. Deletion of YDL193w resulted in a lethal phenotype which could not be suppressed by supplementation with ergosterol or fatty acids. Other deletion mutants were viable under standard conditions, Strains with YBR177c, YMR313c, and YKL140w deleted exhibited phospholipid and/or neutral lipid patterns that were different from the wild-type strain and thus may be further candidate ORFs involved in yeast lipid metabolism.