Milla Paola, Athenstaedt Karin, Viola Franca, Oliaro-Bosso Simonetta, Kohlwein Sepp D, Daum Gunther, Balliano Gianni
Dipartimento di Scienza e Tecnologia del Farmaco, Facoltà di Farmacia, Università degli Studi di Torino, Corso Raffaello 31, I-10125 Torino, Italy.
J Biol Chem. 2002 Jan 25;277(4):2406-12. doi: 10.1074/jbc.M104195200. Epub 2001 Nov 12.
Oxidosqualene cyclase of the yeast encoded by the ERG7 gene converts oxidosqualene to lanosterol, the first cyclic component of sterol biosynthesis. In a previous study (Athenstaedt, K., Zweytick, D., Jandrositz, A, Kohlwein, S. D., and Daum, G. (1999) J. Bacteriol. 181, 6441-6448), Erg7p was identified as a component of yeast lipid particles. Here, we present evidence that Erg7p is almost exclusively associated with this compartment as shown by analysis of enzymatic activity, Western blot analysis, and in vivo localization of Erg7p-GFP. Occurrence of oxidosqualene cyclase in other organelles including the endoplasmic reticulum is negligible. In an erg7 deletion strain or in wild-type cells treated with an inhibitor of oxidosqualene cyclase, the substrate of Erg7p, oxidosqualene, accumulated mostly in lipid particles. Storage in lipid particles of this intermediate produced in excess may provide a possibility to exclude this membrane-perturbing component from other organelles. Thus, our data provide evidence that lipid particles are not only a depot for neutral lipids, but also participate in coordinate sterol metabolism and trafficking and serve as a storage site for compounds that may negatively affect membrane integrity.
由ERG7基因编码的酵母氧化角鲨烯环化酶可将氧化角鲨烯转化为羊毛甾醇,羊毛甾醇是甾醇生物合成的首个环状成分。在之前的一项研究中(Athenstaedt, K., Zweytick, D., Jandrositz, A, Kohlwein, S. D., and Daum, G. (1999) J. Bacteriol. 181, 6441 - 6448),Erg7p被鉴定为酵母脂质颗粒的一个成分。在此,我们提供证据表明,通过对酶活性的分析、蛋白质免疫印迹分析以及Erg7p - GFP的体内定位显示,Erg7p几乎只与该区室相关联。氧化角鲨烯环化酶在包括内质网在内的其他细胞器中的存在可忽略不计。在erg7缺失菌株或用氧化角鲨烯环化酶抑制剂处理的野生型细胞中,Erg7p的底物氧化角鲨烯大多积累在脂质颗粒中。将过量产生的这种中间产物储存在脂质颗粒中,可能提供了一种将这种干扰膜的成分与其他细胞器隔离开的可能性。因此,我们的数据提供了证据表明脂质颗粒不仅是中性脂质的储存库,还参与甾醇的协调代谢和运输,并作为可能对膜完整性产生负面影响的化合物的储存位点。