Taylor F R, Parks L W
J Bacteriol. 1978 Nov;136(2):531-7. doi: 10.1128/jb.136.2.531-537.1978.
The interconversion of free and esterified sterols was followed radioisotopically with [U-14C]acetate and [methyl-14C]methionine. In pulse-chase experiments, radioactivity first appeared mainly in unesterified sterols in exponential-phase cells. Within one generation time, the label equilibrated between the free and esterified sterol pools and subsequently accumulated in steryl esters in stationary-phase cells. When the sterol pools were prelabeled by growing cells aerobically to the stationary phase and the cells were diluted into unlabeled medium, the prelabeled steryl esters returned to the free sterol form under several conditions. (i) During aerobic growth, the prelabeled sterols decreased from 80% to 45% esters in the early exponential phase and then returned to 80% esters as the culture reached the stationary phase. (ii) Under anaerobic conditions, the percentage of prelabeled steryl esters declined continuously. When growth stopped, only 15% of the sterols remained esterified. (iii) In the presence of an inhibitor of sterol biosynthesis, which causes accumulation of a precursor to ergosterol, prelabeled sterols decreased to 40% steryl esters while the precursor was found preferentially in the esterified form. These results indicate that the bulk of the free sterol and steryl ester pools are freely interconvertible, with the steryl esters serving as a supply of free sterols. Furthermore, there is an active cellular control over what types of sterol are found in the free and esterified sterol pools.
利用[U-14C]乙酸盐和[甲基-14C]甲硫氨酸通过放射性同位素追踪法研究了游离甾醇和酯化甾醇的相互转化。在脉冲追踪实验中,放射性最初主要出现在指数生长期细胞的未酯化甾醇中。在一个世代时间内,标记物在游离甾醇库和酯化甾醇库之间达到平衡,随后在稳定期细胞的甾醇酯中积累。当通过好氧培养使细胞生长至稳定期对甾醇库进行预标记,然后将细胞稀释到未标记的培养基中时,在几种条件下预标记的甾醇酯会转化回游离甾醇形式。(i) 在好氧生长过程中,预标记的甾醇在指数生长期早期从80%的酯形式降至45%,然后随着培养物进入稳定期又回到80%的酯形式。(ii) 在厌氧条件下,预标记的甾醇酯百分比持续下降。当生长停止时,只有15%的甾醇仍以酯化形式存在。(iii) 在存在甾醇生物合成抑制剂的情况下,该抑制剂会导致麦角甾醇前体积累,预标记的甾醇降至40%的甾醇酯形式,而前体优先以酯化形式被发现。这些结果表明,大部分游离甾醇库和甾醇酯库可自由相互转化,甾醇酯作为游离甾醇的供应源。此外,细胞对游离甾醇库和酯化甾醇库中存在的甾醇类型有积极的控制作用。