Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.
FEBS J. 2013 Dec;280(23):6033-44. doi: 10.1111/febs.12522. Epub 2013 Oct 11.
We have studied oscillating glycolysis in the strain BY4743 and isogenic strains with deletions of genes encoding enzymes in glycolysis, mitochondrial electron transport and ATP synthesis. We found that deletion of the gene encoding the hexokinase 1 isoform does not affect the oscillations while deletion of the gene encoding the hexokinase 2 isoform results in oscillations with smaller amplitude. The latter is associated with an almost 50% decrease in hexokinase activity. Deletions in the genes encoding the α- and β-subunits of phosphofructokinase abolish the oscillations entirely. This loss in oscillatory activity is associated with a fourfold decrease in phosphofructokinase activity. Deletions of genes encoding subunits of the F1F0 ATPase also inhibit the oscillations in accordance with earlier studies using for example inhibitors. Finally, we identified an apparently new control point involving the mitochondrial cytochrome c oxidase. The latter is difficult to explain as oscillatory activity entails 100% inhibition of this enzyme. The mitochondria of this strain seem to have normal F1F0 ATPase activity. Overall these results support earlier experimental and model studies suggesting that in addition to processes within glycolysis also processes outside this pathway contribute to the control of the oscillatory behaviour.
我们研究了 BY4743 菌株及其糖酵解、线粒体电子传递和 ATP 合成途径中酶缺失的同基因系的振荡糖酵解。我们发现,缺失编码己糖激酶 1 同工酶的基因不会影响振荡,而缺失编码己糖激酶 2 同工酶的基因则导致振荡幅度减小。后者与己糖激酶活性几乎降低 50%有关。磷酸果糖激酶的 α 和 β 亚基编码基因的缺失完全消除了振荡。这种振荡活性的丧失与磷酸果糖激酶活性降低四倍有关。编码 F1F0 ATP 酶亚基的基因缺失也会抑制振荡,这与早期使用抑制剂等的研究一致。最后,我们确定了一个涉及线粒体细胞色素 c 氧化酶的明显新的控制点。后者很难解释,因为振荡活性需要该酶 100%的抑制。该菌株的线粒体似乎具有正常的 F1F0 ATP 酶活性。总的来说,这些结果支持了早期的实验和模型研究,表明除了糖酵解过程中的过程外,该途径之外的过程也有助于控制振荡行为。