Veiga Alexandra, Arrabaça João D, Sansonetty Filipe, Ludovico Paula, Côrte-Real Manuela, Loureiro-Dias Maria C
Laboratório de Microbiologia, Dep. Botânica e Engenharia Biológica, Instituto Superior de Agronomia, 1349-017 Lisboa, Portugal.
FEMS Yeast Res. 2003 Apr;3(2):141-8. doi: 10.1016/S1567-1356(02)00189-7.
Cyanide-resistant respiration (CRR) is a widespread metabolic pathway among yeasts, that involves a mitochondrial alternative oxidase sensitive to salicylhydroxamic acid (SHAM). The physiological role of this pathway has been obscure. We used the yeasts Debaryomyces hansenii and Pichia membranifaciens to elucidate the involvement of CRR in energy conversion. In both yeasts the adenosine triphosphate (ATP) content was still high in the presence of antimycin A or SHAM, but decreased to low levels when both inhibitors were present simultaneously, indicating that CRR was involved in ATP formation. Also the mitochondrial membrane potential (Delta Psi(m)), monitored by fluorescent dyes, was relatively high in the presence of antimycin A and decreased upon addition of SHAM. In both yeasts the presence of complex I was confirmed by the inhibition of oxygen consumption in isolated mitochondria by rotenone. Comparing in the literature the occurrence of CRR and of complex I among yeasts, we found that CRR and complex I were simultaneously present in 12 out of 13 yeasts, whereas in six out of eight yeasts in which CRR was absent, complex I was also absent. Since three phosphorylating sites are active in the main respiratory chain and only one in CRR, we propose a role for this pathway in the fine adjustment of energy provision to the cell.
抗氰呼吸(CRR)是酵母中广泛存在的一种代谢途径,它涉及一种对水杨基羟肟酸(SHAM)敏感的线粒体交替氧化酶。该途径的生理作用一直不清楚。我们使用汉逊德巴利酵母和膜醭毕赤酵母来阐明CRR在能量转换中的作用。在这两种酵母中,存在抗霉素A或SHAM时三磷酸腺苷(ATP)含量仍然很高,但当两种抑制剂同时存在时,ATP含量降至低水平,这表明CRR参与了ATP的形成。同样,通过荧光染料监测的线粒体膜电位(ΔΨm)在存在抗霉素A时相对较高,加入SHAM后降低。在这两种酵母中,通过鱼藤酮对分离线粒体中氧气消耗的抑制作用证实了复合体I的存在。比较文献中酵母中CRR和复合体I的出现情况,我们发现13种酵母中有12种同时存在CRR和复合体I,而在8种不存在CRR的酵母中,有6种也不存在复合体I。由于在主呼吸链中有三个磷酸化位点是活跃的,而在CRR中只有一个,我们提出该途径在对细胞能量供应的精细调节中起作用。