Protein Kinase C Epsilon Promotes Cerebral Ischemic Tolerance Via Modulation of Mitochondrial Sirt5

Sirtuin 5 (SIRT5) is a mitochondrial-localized NAD(+)-dependent lysine desuccinylase and a major regulator of the mitochondrial succinylome. We wanted to determine whether SIRT5 is activated by protein kinase C epsilon (PKC epsilon)-mediated increases in mitochondrial Nampt and whether SIRT5 regulates mitochondrial bioenergetics and neuroprotection against cerebral ischemia. In isolated mitochondria from rat cortical cultures, PKC epsilon activation increased SIRT5 levels and desuccinylation activity in a Nampt-dependent manner. PKC epsilon activation did not lead to significant modifications in SIRT3 activity, the major mitochondrial lysine deacetylase. Assessments of mitochondrial bioenergetics in the cortex of wild type (WT) and SIRT5-/-mice revealed that SIRT5 regulates oxygen consumption in the presence of complex I, complex II, and complex IV substrates. To explore the potential role of SIRT5 in PKC epsilon-mediated protection, we compared WT and SIRT5-/-mice by employing both in vitro and in vivo ischemia paradigms. PKC epsilon-mediated decreases in cell death following oxygen-glucose deprivation were abolished in cortical cultures harvested from SIRT5-/-mice. Furthermore, PKC epsilon failed to prevent cortical degeneration following MCAO in SIRT5-/-mice. Collectively this demonstrates that SIRT5 is an important mitochondrial enzyme for protection against metabolic and ischemic stress following PKC epsilon activation in the brain.