Nes W D, Janssen G G, Crumley F G, Kalinowska M, Akihisa T
Microbial Products Research Unit, Richard B. Russell Research Center, Athens, Georgia 30613.
Arch Biochem Biophys. 1993 Feb 1;300(2):724-33. doi: 10.1006/abbi.1993.1100.
Cultures of Saccharomyces cerevisiae strain GL7 auxotrophic for sterol were incubated with a series of sterols and sterol-like molecules (tetracyclic and pentacyclic triterpenoids) in order to determine the structural requirements of sterols for bulk membrane function. For growth support, the 3 beta-OH group could not be replaced by H, OMe, OBu, NH2, NHOH, OAc, keto, or 3 alpha-OH. A methyl group at C-14 was neither deleterious nor essential for activity. Removal of the C-4 geminal methyl group was obligatory for activity. Thus, no sterol-like triterpenoid supported growth (e.g., tetrahymanol, lanosterol, and cycloartenol). Growth support required a sterol with the longest methylene segment extending from C-20 not to exceed six contiguous C-atoms and the stereochemistry must be C-20 R. No significance could be attributed to branching at C-20 (i.e., to C-21), C-24 (when alkylated), or C-25 (regarding the isopropyl group). Double bonds in the nucleus were not essential for activity since cholestanol supported growth. In several incubations, the addition of trace levels of dietary ergosterol (0.5 microgram/ml) to the medium was necessary to promote growth and transformation of the bulk sterol to a membrane competent sterol(s).
将对甾醇营养缺陷型的酿酒酵母菌株GL7的培养物与一系列甾醇和类甾醇分子(四环和五环三萜类化合物)一起孵育,以确定甾醇对于细胞膜整体功能的结构要求。为了支持生长,3β-OH基团不能被H、OMe、OBu、NH2、NHOH、OAc、酮基或3α-OH取代。C-14位的甲基对活性既无害也非必需。去除C-4偕二甲基是活性所必需的。因此,没有类甾醇三萜类化合物支持生长(例如,四膜虫醇、羊毛甾醇和环阿屯醇)。支持生长需要一种甾醇,其从C-20延伸的最长亚甲基链段不超过六个连续的C原子,并且立体化学必须是C-20 R。C-20(即C-21)、C-24(烷基化时)或C-25(关于异丙基)处的支链没有显著意义。核中的双键对活性不是必需的,因为胆甾烷醇支持生长。在几次孵育中,向培养基中添加微量的膳食麦角甾醇(0.5微克/毫升)对于促进生长以及将整体甾醇转化为具有膜活性的甾醇是必要的。