Structural basis of apoptosis induction by the mitochondrial voltage dependent anion channel

Daniilidis, M.; Guensel, U.; Broutzakis, G.; Janowski, R.; Leitl, K.; Fredriksson, K.; Niessing, D.; Gatsogiannis, C.; Hagn, F. X.

doi:10.1101/2025.04.15.648943

bioRxiv · 10.1101/2025.04.15.648943

Structural basis of apoptosis induction by the mitochondrial voltage dependent anion channel

Daniilidis, M.·Guensel, U.·Broutzakis, G.·Janowski, R.·Leitl, K.·Fredriksson, K.·Niessing, D.·Gatsogiannis, C.·Hagn, F. X.

View original source ↗Export metadata ↓

Abstract

The voltage-dependent anion channel (VDAC) is the main gateway for metabolites across the mitochondrial outer membrane1. In addition, VDAC oligomers have been associated with apoptosis at mitochondrial stress conditions2. However, the mechanistic and structural basis of VDACs capability to induce apoptosis pathways remains poorly understood. Here, we show with biochemical and structural methods that VDAC1 oligomerization triggers the dissociation of its N-terminal -helix (VDAC1-N) from the channel interior. We used advanced lipid nanodiscs as a tool to selectively trap VDAC1 in its canonical helix-inserted and helix-exposed state to facilitate a structural characterization of both conformations by cryo-electron microscopy. The results show that slight changes in the shape and dynamics of the VDAC1 {beta}-barrel suffice to release the N-terminal helix to the channel exterior. This conformational switch addresses the long-standing question how VDAC1 can regulate partner protein binding. To confirm this hypothesis, we performed interaction studies between VDAC1 in both conformational states and the anti-apoptotic partner protein BclxL using nuclear magnetic resonance spectroscopy and could detect binding only for the helix-exposed state. These insights enabled the X-ray structure determination of the BclxL-VDAC1-N complex at high resolution and provided atomistic details on the VDAC1-N binding mode at the BH3-groove in BclxL. Further biochemical assays showed that VDAC1-N promotes pore formation of the pro-apoptotic Bcl2 protein Bak by neutralizing BclxLs inhibitory activity. These findings suggest that stress-induced oligomerization of VDAC can trigger the exposure of its N-terminal -helix leading to the neutralization of anti-apoptotic Bcl2 proteins. This mode-of-action is reminiscent of BH3-only sensitizer Bcl2 proteins3 that are efficient inducers of Bax/Bak-mediated mitochondrial outer membrane permeabilization and ultimately apoptosis.

Source connections

Published article ↗

Explore related subjects

biophysics Preprint

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

Daniilidis, M., Guensel, U., Broutzakis, G., Janowski, R., Leitl, K., Fredriksson, K., Niessing, D., Gatsogiannis, C., Hagn, F. X.. 2025-04-17. Structural basis of apoptosis induction by the mitochondrial voltage dependent anion channel. https://doi.org/10.1101/2025.04.15.648943

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

bioRxiv2026

Design of colipids with controlled lateral distance of head group moieties for targeting strategies in the field of lipid-based nanoparticles

Lipid nanoparticles and liposomes are nanoparticle drug formulations which are firmly established in clinics. In order to expand their application and improve efficacy, surface modifications can be a key strategy, especially for cell specific targeting. Targeting can be achieved by integrating ligand functionalized lipids or lipidoids on the nanoparticle surface. In this case, not solely the targeting ligand itself, but also the structure of the whole targeting lipid/lipidoid can affect the interaction of the targeting ligand with the binding motive on cells. Especially, distal spacer structures between lipid anchor and ligand unit, often composed of polyethylene glycol chains, provide a certain distance of the ligand from the nanoparticle surface. A concept which is currently neglected is the control of the lateral distance between ligand structures on the nanoparticle. Herein we investigated the possibility to design lipidoids with a lateral spacer composed of polyethylene glycol units. We synthesized and characterized different lipidoids which provide a primary amine in the head group for possible ligand coupling. In our model lipidoids the head groups were functionalized with a paramagnetic probe to apply electron paramagnetic resonance (EPR) methods for the characterization of the lipidoid interaction with model nanoparticles, in our case liposomes. Continuous wave EPR was used to study the efficiency of the novel lipidoids to integrate into lipid bilayers of model liposomes, which was shown to depend on lipidoid structure and liposome composition. Furthermore, using double electron-electron resonance (DEER), we observed lipidoid conformational changes occurring upon integration into liposomes.

biophysics↗

bioRxiv2026

Binding mechanisms of therapeutic antibodies to human claudin-18.2

Claudin-18.2 (CLDN18.2) is a tight junction component with highly restricted expression in the paracellular space of nonmalignant gastric epithelial cells. It becomes accessible on tumor cell surface during malignant transformation, making CLDN18.2 an attractive target for targeted therapies against gastric cancer. Recently, zolbetuximab has become the first FDA-approved monoclonal antibody that targets CLDN18.2 in advanced gastric cancer, and others like osemitamab have entered phase 3 clinical trials. However, the high-resolution structure of CLDN18.2 is lacking, and how CLDN18.2 is recognized by therapeutic antibodies remains undefined. Here, we report cryo-EM structures of human claudin-18.2 (hCLDN18.2) in complex with the Fab fragments of zolbetuximab and osemitamab, revealing the structure of hCLDN18.2 and its binding mechanism to the therapeutic antibodies. Further analyses reveal that three residues in the first extracellular loop of hCLDN18.2 that are different from human claudin-18.1 (hCLDN18.1) play a critical role in antibody binding. Our data elucidate the structural basis for selective binding of therapeutic antibodies to hCLDN18.2, which should aid rational design of next-generation immunotherapies targeting hCLDN18.2.

biophysics↗

bioRxiv2026

Structural relationships between the SLC12 cotransporter C-terminal domain, universal stress proteins and adenylating enzymes

Several SLC12 cotransporters bind ATP in a cytoplasmic C-terminal domain (CTD). We analysed 16,192 CTD-containing SLC12 sequences and compared 42,845 representative structures to examine the relationship between this nucleotide site, universal stress proteins (USPs) and adenylating enzymes. In phosphorylated NKCC1, D891 and D927 lie near the ATP phosphate chain, while R801 and K889 form a basic phosphate-binding clamp. NKCC1 retains this arrangement, whereas KCC paralogs replace or lose several corresponding groups. In structural comparisons with USP and adenylating-enzyme references, both CTD lobes score between universal stress proteins and EC-annotated domains, and the two lobes form separate lineages that include bacterial transporters. Lobe-specific and nucleotide-frame comparisons place ATP in the corresponding structural region used for -phosphate chemistry by catalytic members of this fold. Published work on UspG provides a precedent for protein-bound AMP in a natural USP. In the examined SLC12 structures and USP models, candidate protein hydroxyls lack an in-line geometry for attack at ATP P. Protein adenylation would require a change in protein or nucleotide geometry, or a different acceptor. These results define a structural basis for testing conditional protein adenylation at SLC12 nucleotide sites. The required evidence is a covalent protein-AMP adduct, identification of its acceptor and measurement of the expected pyrophosphate coproduct, with soluble nucleotide analysis to distinguish accompanying ATP reactions.

biophysics↗