Naja mossambica mossambica Cobra Cardiotoxin Targets Mitochondria to Disrupt Mitochondrial Membrane Structure and Function.

Boris Zhang,Indira Shrivastava,Edward Gasanoff,Ruben Dagda,Feng Li,Zhengyao Chen

doi:10.3390/toxins11030152

Abstract

Cobra venom cardiotoxins (CVCs) can translocate to mitochondria to promote apoptosis by eliciting mitochondrial dysfunction. However, the molecular mechanism(s) by which CVCs are selectively targeted to the mitochondrion to disrupt mitochondrial function remains to be elucidated. By studying cardiotoxin from Naja mossambica mossambica cobra (cardiotoxin VII4), a basic three-fingered S-type cardiotoxin, we hypothesized that cardiotoxin VII4 binds to cardiolipin (CL) in mitochondria to alter mitochondrial structure/function and promote neurotoxicity. By performing confocal analysis, we observed that red-fluorescently tagged cardiotoxin rapidly translocates to mitochondria in mouse primary cortical neurons and in human SH-SY5Y neuroblastoma cells to promote aberrant mitochondrial fragmentation, a decline in oxidative phosphorylation, and decreased energy production. In addition, by employing electron paramagnetic resonance (EPR) and protein nuclear magnetic resonance (1H-NMR) spectroscopy and phosphorescence quenching of erythrosine in model membranes, our compiled biophysical data show that cardiotoxin VII4 binds to anionic CL, but not to zwitterionic phosphatidylcholine (PC), to increase the permeability and formation of non-bilayer structures in CL-enriched membranes that biochemically mimic the outer and inner mitochondrial membranes. Finally, molecular dynamics simulations and in silico docking studies identified CL binding sites in cardiotoxin VII4 and revealed a molecular mechanism by which cardiotoxin VII4 interacts with CL and PC to bind and penetrate mitochondrial membranes.

Highlights

The ability of a variety of aggregated amyloid-like proteins to interact with lipid bilayers in organelles and cell membranes can occur under certain pathological conditions and in neurodegenerative diseases, which can lead to brain degeneration
electron paramagnetic resonance (EPR) and 1 H-NMR spectroscopy and phosphorescence quenching of erythrosine in model membranes, we further show that cardiotoxin VII4 binds to anionic CL, but not to zwitterionic phosphatidylcholine (PC), to induce the formation of non-bilayer structures in CL enriched membranes that model the outer mitochondrial membrane (OMM) and inner mitochondrial membrane (IMM)
Given its similar biophysical properties as amyloid-like proteins known to target mitochondria (e.g., α-synuclein), we hypothesize that cardiotoxin VII4 can promote mitochondrial dysfunction by binding to anionic phospholipids [7,17]

Summary

Introduction

The ability of a variety of aggregated amyloid-like proteins to interact with lipid bilayers in organelles and cell membranes can occur under certain pathological conditions and in neurodegenerative diseases, which can lead to brain degeneration. Similar effects were observed when neuronal cell lines were treated with oligomeric αS [10,12,13] These effects are linked to the ability of αS oligomers to bind with cardiolipin (CL), an abundant phospholipid of the inner mitochondrial membrane (IMM), which results in the perturbation of the IMM structure [9]. Another consequence of the binding of oligomeric αS to CL is the loss of the function of the mitochondrial adenonsine diphosphate/ adenosine triphosphate (ADP/ATP)

Methods

Results

Discussion

Conclusion