NDLI: The Phospholipase Activity of Ammodytoxin, a Prototype Snake Venom β-Neurotoxin, Is Not Obligatory for Cell Internalisation and Translocation to Mitochondria

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The Phospholipase Activity of Ammodytoxin, a Prototype Snake Venom β-Neurotoxin, Is Not Obligatory for Cell Internalisation and Translocation to Mitochondria

Content Provider	MDPI
Author	Adrijan, Ivanušec Jernej, Šribar Peter, Veranič Igor, Križaj
Copyright Year	2022
Description	β-Neurotoxins are secreted phospholipase $A_{2}$ molecules that inhibit transmission in neuromuscular synapses by poisoning the motor neurons. These toxins specifically and rapidly internalise into the nerve endings of motor neurons. Ammodytoxin (Atx) is a prototype β-neurotoxin from the venom of the nose-horned viper (Vipera ammodytes ammodytes). Here, we studied the relevance of the enzymatic activity of Atx in cell internalisation and subsequent intracellular movement using transmission electron microscopy (TEM). We prepared a recombinant, enzymatically inactive mutant of Atx, Atx(D49S), labelled with gold nanoparticles (GNP), and incubated this with PC12 cells, to analyse its localisation by TEM. Atx(D49S)-GNP internalised into the cells. Inside the cells, Atx(D49S)-GNP was detected in different vesicle-like structures, cytosol, endoplasmic reticulum and mitochondria, where it was spotted in the intermembrane space and matrix. Co-localization of fluorescently labelled Atx(D49S) with mitochondria in PC12 cells by confocal fluorescence microscopy confirmed the reliability of results generated using Atx(D49S)-GNP and TEM and allowed us to conclude that the phospholipase activity of Atx is not obligatory for its cell internalisation and translocation into the mitochondrial intermembrane space and matrix.
Starting Page	375
e-ISSN	20726651
DOI	10.3390/toxins14060375
Journal	Toxins
Issue Number	6
Volume Number	14
Language	English
Publisher	MDPI
Publisher Date	2022-05-28
Access Restriction	Open
Subject Keyword	Toxins Β-neurotoxicity Secreted Phospholipase A2 Ammodytoxin Snake Venom Vipera Ammodytes Ammodytes Transmission Electron Microscopy Localisation Translocation
Content Type	Text
Resource Type	Article

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