Hypoxic-induced truncation of voltage-dependent anion channel 1 is mediated by both asparagine endopeptidase and calpain 1 activities.

ABSTRACT: The voltage-dependent anion channel 1 (VDAC1), an outer mitochondria membrane (OMM) protein, serves as a mitochondrial gatekeeper, mediating the transport of nucleotides, Ca2+ and other metabolites across the OMM. VDAC1 also plays a central role in mitochondria-mediated apoptosis by facilitating the release of apoptotic proteins and by association with both pro- and anti-apoptotic proteins. Tumor cells, which are constantly exposed to hypoxic conditions, affect the cell via the transcription factor hypoxia-inducible factor (HIF) that induces transcriptional activity. In cultured cells and in lung cancer patients, hypoxia induces VDAC1 truncation at the C-terminus (VDAC1-?C). However, the molecular mechanisms involved in VDAC1-?C formation are unknown. Here, we show that hypoxia-induced VDAC1-?C formation is inhibited by the Ca2+ chelator BAPTA-AM, by calpain inhibitor-1, by inhibitor of the asparagine endopeptidase (AEP) and by si-RNA targeting HIF1-? or Ca2+-activated protease calpain-1 expression but not that of calpain-2. Finally, VDAC1-?C expressed in bacteria and reconstituted into a planar lipid bilayer exhibited decreased channel conductance relative to the full-length protein, yet retained voltage-dependent conductance. These findings suggest that hypoxia, acting via HIF-1? expression, leads to VDAC1 cleavage involving the activation of calpain 1 and AEP.

SUBMITTER: Pahima H 

PROVIDER: S-EPMC5849177 | biostudies-literature | 2018 Feb

REPOSITORIES: biostudies-literature