Transduction of the Scorpion Toxin Maurocalcine into Cells

Estève, É.; Mabrouk, Kamel; Dupuis, Alain; Smida-Rezgui, Sophia; Altafaj, Xavier; Grünwald, Didier; Platel, Jean‐Claude; Andreotti, Nicolas; Marty, Isabelle; Sabatier, Jean‐Marc; Ronjat, Michel; Waard, Michel De

doi:10.1074/jbc.m412521200

Cited by 63 publications

(25 citation statements)

References 33 publications

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“…Recent studies have shown that the scorpion toxin maurocalcine crosses the plasma membrane, reaches its target channel located in the endoplasmic reticulum, and consequently causes calcium release from intracellular stores (54). It has been suggested that maurocalcine behaves similarly to the well known cell-penetrating peptides (55), such as the human immunodeficiency virus Tat peptide (56), although recent efforts to understand the translocation mechanism of cell-penetrating peptides have revealed greater complexity involving multiple pathways (57)(58)(59).…”

Section: Discussionmentioning

confidence: 99%

Glycosphingolipid-facilitated Membrane Insertion and Internalization of Cobra Cardiotoxin

Wang

Liu

Lee

et al. 2006

Journal of Biological Chemistry

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Cobra cardiotoxins, a family of basic polypeptides having lipidand heparin-binding capacities similar to the cell-penetrating peptides, induce severe tissue necrosis and systolic heart arrest in snakebite victims. Whereas cardiotoxins are specifically retained on the cell surface via heparan sulfate-mediated processes, their lipid binding ability appears to be responsible, at least in part, for cardiotoxin-induced membrane leakage and cell death. Although the exact role of lipids involved in toxin-mediated cytotoxicity remains largely unknown, monoclonal anti-sulfatide antibody O4 has recently been shown to inhibit the action of CTX A3, the major cardiotoxin from Taiwan cobra venom, on cardiomyocytes by preventing cardiotoxin-induced membrane leakage and CTX A3 internalization into mitochondria. Here, we show that anti-sulfatide acts by blocking the binding of CTX A3 to the sulfatides in the plasma membrane to prevent sulfatide-dependent CTX A3 membrane pore formation and internalization. We also describe the crystal structure of a CTX A3-sulfatide complex in a membrane-like environment at 2.3 Å resolution. The unexpected orientation of the sulfatide fatty chains in the structure allows prediction of the mode of toxin insertion into the plasma membrane. CTX A3 recognizes both the headgroup and the ceramide interfacial region of sulfatide to induce a lipid conformational change that may play a key role in CTX A3 oligomerization and cellular internalization. This proposed lipid-mediated toxin translocation mechanism may also shed light on the cellular uptake mechanism of the amphiphilic cell-penetrating peptides known to involve multiple internalization pathways.Glycosphingolipids in eukaryotic cells have important biological functions in membrane trafficking and cell signaling (1, 2). The number of disease-related proteins found to interact specifically with glycosphingolipids is also increasing (3-5). For instance, recent studies have shown that natural killer cells can be activated by glycosylceramides from the cell surface of Gram-negative bacteria that do not contain lipopolysaccharide (6, 7). There is also evidence indicating that, in response to tumor necrosis factor-␣, trafficking of ganglioside GD3 from the plasma membrane to mitochondria can trigger several cell death signaling responses (3,8,9). Although the molecular mechanisms accounting for the observed glycosphingolipid transport and glycolipidtriggered cell responses remain elusive, recent progress in determining the three-dimensional structure of protein⅐glycosphingolipid complexes has shed some light on the process (10 -13). These glycosphingolipid-binding proteins function by either extracting lipids from the membrane and forming a water-soluble protein⅐lipid complex, as in the cases of glycosphingolipid transfer protein, saposin B, and lipid-presenting CD1a (10 -12), or by localizing to a glycosphingolipid domain of the plasma membrane and eventually being internalized via the endocytic pathway, as seen for cholera and Shiga toxins (13,14). Aft...

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Section: Discussionmentioning

confidence: 99%

Glycosphingolipid-facilitated Membrane Insertion and Internalization of Cobra Cardiotoxin

Wang

Liu

Lee

et al. 2006

Journal of Biological Chemistry

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“…As previously (7,9,22), through macropinocytosis (23). The cell penetration of MCa b / Strep complexes involves one or multiple steps of attachment to the membrane through binding to cell surface glycosaminoglycans and negatively charged lipids.…”

Section: Removing Disulfide Bridges In Mca Disrupts the Secondarymentioning

confidence: 99%

Design of a Disulfide-less, Pharmacologically Inert, and Chemically Competent Analog of Maurocalcine for the Efficient Transport of Impermeant Compounds into Cells

Ram

Weiss

Texier

et al. 2008

Journal of Biological Chemistry

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Maurocalcine is a 33-mer peptide initially isolated from the venom of a Tunisian scorpion. It has proved itself valuable as a pharmacological activator of the ryanodine receptor and has helped the understanding of the molecular basis underlying excitation-contraction coupling in skeletal muscles. Because of its positively charged nature, it is also an innovative vector for the cell penetration of various compounds. We report a novel maurocalcine analog with improved properties: (i) the complete loss of pharmacological activity, (ii) preservation of the potent ability to carry cargo molecules into cells, and (iii) coupling chemistries not affected by the presence of internal cysteine residues of maurocalcine. We did this by replacing the six internal cysteine residues of maurocalcine by isosteric 2-aminobutyric acid residues and by adding an additional N-terminal biotinylated lysine (for a proof of concept analog) or an N-terminal cysteine residue (for a chemically competent coupling analogue). Additional replacement of a glutamate residue by alanyl at position 12 further improves the potency of these analogues. Coupling to several cargo molecules or nanoparticles are presented to illustrate the cell penetration potency and usefulness of these pharmacologically inactive analogs. Maurocalcine (MCa)3 is a highly basic 33-mer peptide isolated from the venom of the scorpion Scorpio maurus palmatus. It efficiently binds to the ryanodine receptor of skeletal muscles (RyR1 isoform) (1) and promotes channel opening to promote calcium release from the sarcoplasmic reticulum (SR). This pharmacological effect of MCa can be indirectly monitored through the stimulation it exerts on [ 3 H]ryanodine binding (2). In muscle fibers, MCa produces a transient loss of voltage control of Ca 2ϩ release from RyR1 channels. This effect is due to an alteration of repolarization-induced closure of RyR1 channels, a process normally under the control of voltage-dependent dihydropyridine (DHP)-sensitive calcium channels (3). This function of MCa is due to a partial sequence homology between MCa and a cytoplasmic loop of the DHP-sensitive channel (2). These observations explain why MCa, along with other members of the same family of toxins such as imperatoxin 1A (4), hemicalcin (5), and opicalcin 1 and 2 (6) are useful both for their pharmacological properties and for deciphering fine molecular details of the excitation-contraction coupling process.Recently, MCa has also proven of interest for its property of efficiently crossing the plasma membrane, either alone or when coupled to a membrane-impermeant cargo protein (2, 7). MCa is a highly charged peptide with 12 basic residues out of 33, and a net global positive charge of ϩ8. Most of these residues are on one face of the molecule, the opposite face being mostly hydrophobic in nature. The rich content of basic amino acid residues of MCa is reminiscent of that of all cell-penetrating peptides (CPPs) characterized so far (Tat, penetratin, and poly-R). Hence, MCa can be classified within an eme...

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“…These observations suggested that MCa is able to cross the plasma membrane to reach its pharmacological target. This was first demonstrated when a biotinylated analogue of MCa was coupled to a fluorescent derivative of streptavidin, and the complex was shown to cross the plasma membrane (4). Cell penetration of this MCa-based complex is rapid, reaches saturation within minutes, and occurs at concentrations as low as 10 nM (5).…”

Section: Maurocalcine (Mca)mentioning

confidence: 99%

Direct Peptide Interaction with Surface Glycosaminoglycans Contributes to the Cell Penetration of Maurocalcine

Ram

Aroui

Jaumain

et al. 2008

Journal of Biological Chemistry

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Transduction of the Scorpion Toxin Maurocalcine into Cells

Cited by 63 publications

References 33 publications

Glycosphingolipid-facilitated Membrane Insertion and Internalization of Cobra Cardiotoxin

Glycosphingolipid-facilitated Membrane Insertion and Internalization of Cobra Cardiotoxin

Design of a Disulfide-less, Pharmacologically Inert, and Chemically Competent Analog of Maurocalcine for the Efficient Transport of Impermeant Compounds into Cells

Direct Peptide Interaction with Surface Glycosaminoglycans Contributes to the Cell Penetration of Maurocalcine

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