Avery S V, Smith S L, Ghazi A M, Hoptroff M J
Department of Biology, University Plaza, Georgia State University, Atlanta, Georgia 30302-4010, USA.
Appl Environ Microbiol. 1999 Mar;65(3):1191-7. doi: 10.1128/AEM.65.3.1191-1197.1999.
The influence of modified plasma membrane fatty acid composition on cellular strontium accumulation in Saccharomyces cerevisiae was investigated. Growth of S. cerevisiae in the presence of 1 mM linoleate (18:2) (which results in 18:2 incorporation to approximately 70% of total cellular and plasma membrane fatty acids, with no effect on growth rate) yielded cells that accumulated Sr2+ intracellularly at approximately twice the rate of S. cerevisiae grown without a fatty acid supplement. This effect was evident over a wide range of external Sr2+ concentrations (25 microM to 5 mM) and increased with the extent of cellular 18:2 incorporation. Stimulation of Sr2+ accumulation was not evident following enrichment of S. cerevisiae with either palmitoleate (16:1), linolenate (18:3) (n-3 and n-6 isomers), or eicosadienoate (20:2) (n-6 and n-9 isomers). Competition experiments revealed that Ca2+- and Mg2+-induced inhibition of Sr2+ accumulation did not differ between unsupplemented and 18:2-supplemented cells. Treatment with trifluoperazine (TFP) (which can act as a calmodulin antagonist and Ca2+-ATPase inhibitor), at a low concentration that precluded nonspecific K+ efflux, increased intracellular Sr2+ accumulation by approximately 3.6- and 1.4-fold in unsupplemented and 18:2-supplemented cells, respectively. Thus, TFP abolished the enhanced Sr2+ accumulation ability of 18:2-supplemented cells. Moreover, the rate of Sr2+ release from Sr2+-loaded fatty acid-unsupplemented cells was found to be at least twice as great as that from Sr2+-loaded 18:2-enriched cells. The influence of enrichment with other fatty acids on Sr2+ efflux was variable. The results reveal an enhanced Sr2+ accumulation ability of S. cerevisiae following 18:2-enrichment, which is attributed to diminished Sr2+ efflux activity in these cells.
研究了修饰的质膜脂肪酸组成对酿酒酵母细胞锶积累的影响。酿酒酵母在1 mM亚油酸(18:2)存在下生长(这导致18:2掺入到约70%的总细胞脂肪酸和质膜脂肪酸中,对生长速率无影响),产生的细胞在细胞内积累Sr2+的速率约为未添加脂肪酸生长的酿酒酵母的两倍。在很宽的外部Sr2+浓度范围(25 microM至5 mM)内这种效应都很明显,并且随着细胞18:2掺入程度的增加而增强。用棕榈油酸(16:1)、亚麻酸(18:3)(n-3和n-6异构体)或二十碳二烯酸(20:2)(n-6和n-9异构体)富集酿酒酵母后,未观察到对Sr2+积累的刺激作用。竞争实验表明,未添加脂肪酸和添加18:2的细胞之间,Ca2+和Mg2+对Sr2+积累的抑制作用没有差异。用三氟拉嗪(TFP)(可作为钙调蛋白拮抗剂和Ca2+-ATP酶抑制剂)处理,在低浓度下排除了非特异性K+外流,未添加脂肪酸和添加18:2的细胞内Sr2+积累分别增加了约3.6倍和1.4倍。因此,TFP消除了添加18:2的细胞增强的Sr2+积累能力。此外,发现从负载Sr2+的未添加脂肪酸的细胞中Sr2+释放速率至少是从负载Sr2+的富含18:2的细胞中Sr2+释放速率的两倍。用其他脂肪酸富集对Sr2+外流的影响是可变的。结果表明,18:2富集后酿酒酵母的Sr2+积累能力增强,这归因于这些细胞中Sr2+外流活性的降低。