Shuvo 2019 J Bioenerg Biomembr
| Shuvo SR, Wiens LM, Subramaniam S, Treberg JR, Court DA (2019) Increased reactive oxygen species production and maintenance of membrane potential in VDAC-less Neurospora crassa mitochondria. J Bioenerg Biomembr 51:341-54. |
Shuvo SR, Wiens LM, Subramaniam S, Treberg JR, Court DA (2019) J Bioenerg Biomembr
Abstract: The highly abundant voltage-dependent anion-selective channel (VDAC) allows transit of metabolites across the mitochondrial outer membrane. Previous studies in Neurospora crassa showed that the LoPo strain, expressing 50% of normal VDAC levels, is indistinguishable from wild-type (WT). In contrast, the absence of VDAC (ΞPor-1), or the expression of an N-terminally truncated variant VDAC (ΞN2-12porin), is associated with deficiencies in cytochromes b and aa3 of complexes III and IV and concomitantly increased alternative oxidase (AOX) activity. These observations led us to investigate complex I and complex II activities in these strains, and to explore their mitochondrial bioenergetics. The current study reveals that the total NADH dehydrogenase activity is similar in mitochondria from WT, LoPo, ΞPor-1 and ΞN2-12porin strains; however, in ΞPor-1 most of this activity is the product of rotenone-insensitive alternative NADH dehydrogenases. Unexpectedly, LoPo mitochondria have increased complex II activity. In all mitochondrial types analyzed, oxygen consumption is higher in the presence of the complex II substrate succinate, than with the NADH-linked (complex I) substrates glutamate and malate. When driven by a combination of complex I and II substrates, membrane potentials (ΞΟ) and oxygen consumption rates (OCR) under non-phosphorylating conditions are similar in all mitochondria. However, as expected, the induction of state 3 (phosphorylating) conditions in ΞPor-1 mitochondria is associated with smaller but significant increases in OCR and smaller decreases in ΞΟ than those seen in wild-type mitochondria. High ROS production, particularly in the presence of rotenone, was observed under non-phosphorylating conditions in the ΞPor-1 mitochondria. Thus, the absence of VDAC is associated with increased ROS production, in spite of AOX activity and wild-type OCR in ΞPor-1 mitochondria. β’ Keywords: Alternative oxidase, Membrane potential, Mitochondrial porin, Neurospora crassa, Reactive oxygen species, VDAC β’ Bioblast editor: Plangger M β’ O2k-Network Lab: CA Winnipeg Treberg JR
Labels: MiParea: Respiration, mt-Membrane
Organism: Fungi
Preparation: Isolated mitochondria Enzyme: Complex I, Complex II;succinate dehydrogenase Regulation: mt-Membrane potential Coupling state: LEAK, OXPHOS, ET Pathway: N, S, NS, ROX HRR: Oxygraph-2k
Labels, 2019-08
