Bligny R, Rebeillé F, Douce R
J Biol Chem. 1985 Aug 5;260(16):9166-70.
Sycamore cells (Acer pseudoplatanus L.) in suspension culture were grown at 25 degrees C in culture medium containing two oxygen concentrations: 250 microM O2 (standard conditions) and 10 microM O2 (O2-limiting conditions). The decrease of O2 concentration in the culture medium did not modify significantly the relative proportion of each phospholipid. In contrast, the molar proportion of fatty acids was dramatically changed in all lipid classes of the cell membranes; the average percentage of oleate increased from 3 to 45% whereas that of linoleate decreased from 49 to 22%. When normal culture conditions were restored (250 microM O2), oleate underwent a rapid desaturation process; the loss of oleic acid was associated with a stoichiometric appearance of linoleic acid at a rate of about 4 nmol of oleate desaturated/h/10(6) cells. Under these conditions, no change in the Arrhenius-type plots of the rate of sycamore cell respiration was observed; the values of the transition temperature and of the Arrhenius activation energy (Ea) associated with the cell respiration as well as with the respiration-associated enzymes remained unchanged. Thus it was concluded that the fact that a strong decrease in the fraction of unsaturated fatty acid residues present in the mitochondria had no effect on electron transport rates and Arrhenius plot discontinuities casts doubt on the significance of such changes in terms of chilling injury. Finally it is suggested that some of the Arrhenius discontinuities observed at the level of membrane enzyme could be the consequence of intrinsic thermotropic changes in protein arrangement independent of lipid fluidity.
悬铃木细胞(槭树)在悬浮培养中于25℃下在含有两种氧浓度的培养基中生长:250微摩尔O₂(标准条件)和10微摩尔O₂(O₂限制条件)。培养基中氧浓度的降低并未显著改变各磷脂的相对比例。相反,细胞膜所有脂质类别的脂肪酸摩尔比例发生了显著变化;油酸的平均百分比从3%增加到45%,而亚油酸的平均百分比从49%下降到22%。当恢复正常培养条件(250微摩尔O₂)时,油酸经历了快速去饱和过程;油酸的损失与亚油酸的化学计量出现相关,去饱和速率约为4纳摩尔油酸/小时/10⁶个细胞。在这些条件下,未观察到悬铃木细胞呼吸速率的阿累尼乌斯型图有变化;与细胞呼吸以及与呼吸相关酶相关的转变温度和阿累尼乌斯活化能(Ea)值保持不变。因此得出结论,线粒体中不饱和脂肪酸残基比例的大幅下降对电子传递速率和阿累尼乌斯图不连续性没有影响,这使人对这种变化在冷害方面的意义产生怀疑。最后有人提出,在膜酶水平观察到的一些阿累尼乌斯不连续性可能是蛋白质排列内在热致变化的结果,与脂质流动性无关。