Poneratoxin is a paralyzing

neurotoxic Neurotoxicity is a form of toxicity in which a biological, chemical, or physical agent produces an adverse effect on the structure or function of the central and/or peripheral nervous system. It occurs when exposure to a substance – specificall ...

peptide Peptides (, ) are short chains of amino acids linked by peptide bonds. Long chains of amino acids are called proteins. Chains of fewer than twenty amino acids are called oligopeptides, and include dipeptides, tripeptides, and tetrapeptides. A ...

made by the bullet ant ''

Paraponera clavata ''Paraponera clavata'' is a species of ant, commonly known as the bullet ant, named for its extremely painful sting. It inhabits humid lowland rainforests in Central and South America. Etymology The specific epithet of the ant, ', means "club- ...

''. It prevents inactivation of

voltage gated sodium channels Sodium channels are integral membrane proteins that form ion channels, conducting sodium ions (Na+) through a cell's membrane. They belong to the superfamily of cation channels and can be classified according to the trigger that opens the channel ...

and therefore blocks the

synaptic transmission Neurotransmission (Latin: ''transmissio'' "passage, crossing" from ''transmittere'' "send, let through") is the process by which signaling molecules called neurotransmitters are released by the axon terminal of a neuron (the presynaptic neuron), ...

in the

central nervous system The central nervous system (CNS) is the part of the nervous system consisting primarily of the brain and spinal cord. The CNS is so named because the brain integrates the received information and coordinates and influences the activity of all par ...

. Specifically, poneratoxin acts on voltage gated sodium channels in

skeletal muscle Skeletal muscles (commonly referred to as muscles) are organs of the vertebrate muscular system and typically are attached by tendons to bones of a skeleton. The muscle cells of skeletal muscles are much longer than in the other types of muscl ...

fibers, causing paralysis, and

nociceptive Nociception (also nocioception, from Latin ''nocere'' 'to harm or hurt') is the Somatosensory system, sensory nervous system's process of encoding Noxious stimulus, noxious stimuli. It deals with a series of events and processes required for an org ...

fibers, causing pain. It is rated as a 4 plus on the

Schmidt sting pain index The Schmidt sting pain index is a pain scale rating the relative pain caused by different hymenopteran stings. It is mainly the work of Justin O. Schmidt (born 1947), an entomologist at the Carl Hayden Bee Research Center in Arizona, United States ...

, the highest possible rating with that system, and its effects can cause waves of pain up to twelve hours after a single sting. Schmidt describes it as "pure, intense, brilliant pain...like walking over flaming charcoal with a three-inch nail embedded in your heel." It is additionally being studied for its uses in biological insecticides.

Mechanism of action

Overall, poneratoxin disrupts normal function of

voltage-gated sodium channels Sodium channels are integral membrane proteins that form ion channels, conducting sodium ions (Na+) through a cell's membrane. They belong to the superfamily of cation channels and can be classified according to the trigger that opens the channel ...

in both

vertebrate Vertebrates () comprise all animal taxa within the subphylum Vertebrata () ( chordates with backbones), including all mammals, birds, reptiles, amphibians, and fish. Vertebrates represent the overwhelming majority of the phylum Chordata, ...

s and

invertebrate Invertebrates are a paraphyletic group of animals that neither possess nor develop a vertebral column (commonly known as a ''backbone'' or ''spine''), derived from the notochord. This is a grouping including all animals apart from the chordate ...

s. It causes repetitive firing and prolongation of

action potential An action potential occurs when the membrane potential of a specific cell location rapidly rises and falls. This depolarization then causes adjacent locations to similarly depolarize. Action potentials occur in several types of animal cells, ...

s, particularly in the

of insects. The increase in signaling is what causes the intense pain from

bullet ant A bullet is a kinetic projectile, a component of firearm ammunition that is shot from a gun barrel. Bullets are made of a variety of materials, such as copper, lead, steel, polymer, rubber and even wax. Bullets are made in various shapes and c ...

stings. Poneratoxin is in an inactive state when stored in the ant venom reservoir due to the reservoir's acidic conditions, but it becomes toxic when activated via a multistep process. The combination of poneratoxin binding to a

cell membrane The cell membrane (also known as the plasma membrane (PM) or cytoplasmic membrane, and historically referred to as the plasmalemma) is a biological membrane that separates and protects the interior of all cells from the outside environment ( ...

(in order to act upon a voltage-gated sodium channel) and the movement from acidic conditions in the ant venom reservoir to basic conditions at the target site leads to poneratoxin undergoing a

conformational change In biochemistry, a conformational change is a change in the shape of a macromolecule, often induced by environmental factors. A macromolecule is usually flexible and dynamic. Its shape can change in response to changes in its environment or oth ...

that activates it. Catterall ''et al.'' hypothesized that some

polypeptide Peptides (, ) are short chains of amino acids linked by peptide bonds. Long chains of amino acids are called proteins. Chains of fewer than twenty amino acids are called oligopeptides, and include dipeptides, tripeptides, and tetrapeptides. A p ...

neurotoxin Neurotoxins are toxins that are destructive to nerve tissue (causing neurotoxicity). Neurotoxins are an extensive class of exogenous chemical neurological insultsSpencer 2000 that can adversely affect function in both developing and mature ner ...

s that modify voltage-gated channels function via a "voltage-sensor trapping" mechanism. The hypothesis states that neurotoxins similar to poneratoxin, such as alpha-scorpion toxins, act upon

sodium channel Sodium channels are integral membrane proteins that form ion channels, conducting sodium ions (Na+) through a cell's membrane. They belong to the superfamily of cation channels and can be classified according to the trigger that opens the channel ...

s via binding to the channels'

receptor Receptor may refer to: * Sensory receptor, in physiology, any structure which, on receiving environmental stimuli, produces an informative nerve impulse *Receptor (biochemistry), in biochemistry, a protein molecule that receives and responds to a ...

site 3, which normally affects the channels' ability to inactivate. Therefore, receptor site 3 neurotoxins often affect sodium channels by slowing or blocking inactivation. Normally, the region of the channel where neurotoxin receptor site 3 is undergoes a conformational change of an outward movement to lead to inactivation. Receptor site 3 neurotoxins are proposed to prevent this conformational change via interaction with

acid In computer science, ACID ( atomicity, consistency, isolation, durability) is a set of properties of database transactions intended to guarantee data validity despite errors, power failures, and other mishaps. In the context of databases, a sequ ...

ic and

hydrophobic In chemistry, hydrophobicity is the physical property of a molecule that is seemingly repelled from a mass of water (known as a hydrophobe). In contrast, hydrophiles are attracted to water. Hydrophobic molecules tend to be nonpolar and, th ...

amino acid residues Protein structure is the three-dimensional arrangement of atoms in an amino acid-chain molecule. Proteins are polymers specifically polypeptides formed from sequences of amino acids, the monomers of the polymer. A single amino acid monomer may ...

at that site. When frog

skeletal muscle fibers Skeletal muscles (commonly referred to as muscles) are organs of the vertebrate muscular system and typically are attached by tendons to bones of a skeleton. The muscle cells of skeletal muscles are much longer than in the other types of muscle ...

were exposed to poneratoxin, it was found that poneratoxin primarily affected voltage-dependent sodium channels by decreasing the peak sodium current and also inducing a slow sodium current. This combination resulted in the sodium channels activating at very negative potentials and deactivating very slowly, a phenomenon commonly seen in excitable tissues. Poneratoxin is considered as a slow-acting

agonist An agonist is a chemical that activates a receptor to produce a biological response. Receptors are cellular proteins whose activation causes the cell to modify what it is currently doing. In contrast, an antagonist blocks the action of the ago ...

for

smooth muscles Smooth muscle is an involuntary non-striated muscle, so-called because it has no sarcomeres and therefore no striations (''bands'' or ''stripes''). It is divided into two subgroups, single-unit and multiunit smooth muscle. Within single-unit mus ...

Structure

The poneratoxin peptide is stored in an inactive 25-residue peptide (amino acid sequence FLPLLILGSLLMTPPVIQAIHDAQR) in the venom reservoir of ''

''. The

secondary structure Protein secondary structure is the three dimensional conformational isomerism, form of ''local segments'' of proteins. The two most common Protein structure#Secondary structure, secondary structural elements are alpha helix, alpha helices and beta ...

is characterized by a

helix-turn-helix Helix-turn-helix is a DNA-binding protein (DBP). The helix-turn-helix (HTH) is a major structural motif capable of binding DNA. Each monomer incorporates two α helices, joined by a short strand of amino acids, that bind to the major groove of D ...

motif: two

alpha helices The alpha helix (α-helix) is a common motif in the secondary structure of proteins and is a right hand-helix conformation in which every backbone N−H group hydrogen bonds to the backbone C=O group of the amino acid located four residues ear ...

connected by a

beta-turn β turns (also β-bends, tight turns, reverse turns, Venkatachalam turns) are the most common form of turns—a type of non-regular secondary structure in proteins that cause a change in direction of the polypeptide chain. They are very common mot ...

. The two

are formed by residues 3–9 at the N-terminus, and residues 17–24 at the C-terminus, and they are connected by the

at residues 11–16. From a three-dimensional perspective, this structure forms a preferential V-shape with the two helices undergoing loose non-covalent interactions with each other. This is notable because of its structural similarity to other peptides that interact with the membrane, and indicates that poneratoxin will also interact with the membrane and thereby affect imbedded

. Furthermore, the structure of the peptide shifts from a

random coil In polymer chemistry, a random coil is a conformation of polymers where the monomer subunits are oriented randomly while still being bonded to adjacent units. It is not one specific shape, but a statistical distribution of shapes for all the cha ...

to the structured

when introduced to a

lipid bilayer The lipid bilayer (or phospholipid bilayer) is a thin polar membrane made of two layers of lipid molecules. These membranes are flat sheets that form a continuous barrier around all cells. The cell membranes of almost all organisms and many vir ...

environment, which indicates that this motif is important for interacting with the membrane. The two alpha helices, however, have markedly different characteristics. The N-terminal alpha helix is apolar, containing a central

core with

hydrophilic A hydrophile is a molecule or other molecular entity that is attracted to water molecules and tends to be dissolved by water.Liddell, H.G. & Scott, R. (1940). ''A Greek-English Lexicon'' Oxford: Clarendon Press. In contrast, hydrophobes are no ...

residues at either end, and is uncharged. It is similar in structure to a transmembrane

signal peptide A signal peptide (sometimes referred to as signal sequence, targeting signal, localization signal, localization sequence, transit peptide, leader sequence or leader peptide) is a short peptide (usually 16-30 amino acids long) present at the N-ter ...

, which implies that it will anchor at the membrane by burying the hydrophobic core within the bilayer. In particular, the bulky and very hydrophobic

phenylalanine Phenylalanine (symbol Phe or F) is an essential α-amino acid with the formula . It can be viewed as a benzyl group substituted for the methyl group of alanine, or a phenyl group in place of a terminal hydrogen of alanine. This essential amino a ...

residue is important for interacting with uncharged lipid bilayers, such as those composed of

phosphatidylcholine Phosphatidylcholines (PC) are a class of phospholipids that incorporate choline as a headgroup. They are a major component of biological membranes and can be easily obtained from a variety of readily available sources, such as egg yolk or soyb ...

. The C-terminal alpha helix is amphipathic with one side displaying polar and charged residues, and the other displaying non-polar residues, which drives insertion into the plasma membrane. Specifically, the positively charged

arginine Arginine is the amino acid with the formula (H2N)(HN)CN(H)(CH2)3CH(NH2)CO2H. The molecule features a guanidino group appended to a standard amino acid framework. At physiological pH, the carboxylic acid is deprotonated (−CO2−) and both the am ...

and the non-polar

alanine Alanine (symbol Ala or A), or α-alanine, is an α-amino acid that is used in the biosynthesis of proteins. It contains an amine group and a carboxylic acid group, both attached to the central carbon atom which also carries a methyl group side c ...

residues were both shown to be essential for poneratoxin potency. See figure, where the hydrophobic (red) and hydrophilic (blue) regions of poneratoxin and the lipid bilayer align, demonstrating that the structure is evolved to insert into the membrane, which will promote interaction with the

Toxicology

Many people consider a sting from a

to resemble the sensation of getting shot. Justin Schmidt, an

entomologist Entomology () is the scientific study of insects, a branch of zoology. In the past the term "insect" was less specific, and historically the definition of entomology would also include the study of animals in other arthropod groups, such as arach ...

who developed the

, described it as "pure, intense, brilliant pain...like walking over flaming charcoal with a three-inch nail embedded in your heel," and considers the sting from a bullet ant to be the most painful insect sting he has experienced. The pain from bullet ant stings can last for many hours, even up to 24 hours. Both the immense pain and the duration of the sting are due to the effects of poneratoxin. In addition to the notorious pain, symptoms of stings from bullet ants (as well as stings from other ants of the genus '' Paraponera'' as well as the genus ''

Dinoponera ''Dinoponera'' is a strictly South American genus of ant in the subfamily Ponerinae, commonly called tocandiras or giant Amazonian ants. These ants are generally less well known than ''Paraponera clavata'', the bullet ant, yet ''Dinoponera'' f ...

'') include fever, cold sweats, nausea, vomiting,

lymphadenopathy Lymphadenopathy or adenopathy is a disease of the lymph nodes, in which they are abnormal in size or consistency. Lymphadenopathy of an inflammatory type (the most common type) is lymphadenitis, producing swollen or enlarged lymph nodes. In cli ...

and

cardiac arrhythmia Arrhythmias, also known as cardiac arrhythmias, heart arrhythmias, or dysrhythmias, are irregularities in the heartbeat, including when it is too fast or too slow. A resting heart rate that is too fast – above 100 beats per minute in adults ...

s. Toxicity assays have found that the LT₅₀ of poneratoxin, delivered via viral injections to ''S. frugiperda'' larvae, was at 131 hours post-injection. A dose of 10⁵ pfu of poneratoxin was sufficient to kill the ''S. frugiperda'' larvae, and a dose of 10 ng could paralyze them. Based on these experiments, scientists believe poneratoxin can make a good candidate as a bio-

insecticide Insecticides are substances used to kill insects. They include ovicides and larvicides used against insect eggs and larvae, respectively. Insecticides are used in agriculture, medicine, industry and by consumers. Insecticides are claimed to b ...

because of its neurotoxicity to other insects, making it capable of immobilizing or even killing insects infected with it. The making of a

recombinant virus A recombinant virus may occur naturally or be produced by recombining pieces of DNA using recombinant DNA technology. Synthetic recombination This may be used to produce viral vaccines or gene therapy vectors. Natural recombination The term ...

by engineering a

baculovirus ''Baculoviridae'' is a family of viruses. Arthropod Arthropods (, (gen. ποδός)) are invertebrate animals with an exoskeleton, a Segmentation (biology), segmented body, and paired jointed appendages. Arthropods form the phylum Arthropod ...

that expresses poneratoxin has been proposed.

References

External links

Antinociceptive effect of poneratoxin (PoTX) in rats. Pestycydy, 2008, (1-2), 135-141. ISSN 0208-8703
* {{Ion channel modulators Ion channel toxins Neurotoxins Peripheral membrane proteins Myrmecology Sodium channel openers

Mechanism of action

Structure

Toxicology

See also

References

External links