Mitochondrial LON protease-dependent degradation of cytochrome c oxidase subunits under hypoxia and myocardial ischemia

作者：Sepuri, Naresh B. V.; Angireddy, Rajesh; Srinivasan, Satish; Guha, Manti; Spear, Joseph; Lu, Bin; Anandatheerthavarada, Hindupur K.; Suzuki, Carolyn K.; Avadhani, Narayan G.^*

来源：Biochimica et Biophysica Acta-Bioenergetics, 2017, 1858(7): 519-528.

DOI：10.1016/j.bbabio.2017.04.003

• Mitochondrial LON
• CcO subunits
• PKA dependent phosphorylation
• Hypoxia
• Heart ischemia
• 3D modeling

The mitochondrial ATP dependent matrix protease, Lon, is involved in the maintenance of mitochondrial DNA nucleoids and degradation of abnormal or misfolded proteins. The Lon protease regulates mitochondrial Tfam (mitochondrial transcription factor A) level and thus modulates mitochondrial DNA (mtDNA) content. We have previously shown that hypoxic stress induces the PICA-dependent phosphorylation of cytochrome c oxidase (CcO) subunits I, IVil, and Vb and a time-dependent reduction of these subunits in RAW 264.7 murine macrophages subjected to hypoxia and rabbit hearts subjected to ischemia/reperfusion. Here, we show that Lon is involved in the preferential turnover of phosphorylated CcO subunits under hypoxic/ischemic stress. Induction of Lon protease occurs at 6 to 12 h of hypoxia and this increase coincides with lower CcO subunit contents. Over-expression of flag-tagged wild type and phosphorylation site mutant Vb and IVil subunits (S40A and T52A, respectively) caused marked degradation of wild type protein under hypoxia while the mutant proteins were relatively resistant. Furthermore, the recombinant purified Lon protease degraded the phosphorylated IVil and Vb subunits, while the phosphorylation-site mutant proteins were resistant to degradation. 3D structural modeling shows that the phosphorylation sites are exposed to the matrix compartment, accessible to matrix PICA and Lon protease. Hypoxic stress did not alter CcO subunit levels in Lon depleted cells, confirming its role in CcO turnover. Our results therefore suggest that Lon preferentially degrades the phosphorylated subunits of CcO and plays a role in the regulation of CcO activity in hypoxia and ischemia/reperfusion injury.

• 出版日期2017-7
• 单位温州医科大学