Polyamines

Despite the vast majority of spiders being harmless to humans, many spider venoms
contain in their venoms potent small molecules called polyamines. Polyamines are often
very effective and specific blockers of ion channels or of receptors. Their target receptors
are those which recognize excitatory amino acids in the mammalian central nervous
system and are classified into three major subtypes, ones which prefer !methyl!D!
aspartate "#D$%, &uis&ualate "'$%, or kainate "($% as type agonists respectively .
$rgiotoxin, a glutamate!activated postsynaptic ion channel blocking polyamine from the
spider Argiope lobata.
)pider toxins have been isolated which are selective and reversible noncompetitive
antagonists of #D$ "!methyl!D!aspartate% receptors in the mammalian brain. *y way
of example, Dolomedes okefinokensii "+ishing spider% bites rarely cause anything but the
mildest of edema and possibly minor necrosis. The venom, however, has yielded a
compound that may have use as an analgesic, a polyamine that reversibly blocks ,!and
-!type voltage!sensitive calcium channels.


.ther examples of the usefulness of spider polyamines are the alpha!agatoxins $ga!/0
and $ga!/1 from the $merican funnel web spider Agelenopsis aperta. These low
molecular weight molecules cause rapid, reversible paralysis correlated with use!
dependent postsynaptic block of excitatory postsynaptic potential and ionophoretic
glutamate potentials. $ga!/0 is a toxin capable of inhibiting an elusive neuronal calcium
channel that is resistant to blockage by either omega!conotoxin /20$ or nifedipine
making this component a novel tool with which to probe this channel. 0ntravenous or
intracerebroventricular administration of $ga!/1 produces dose!dependent suppression
of behavioral convulsions induced in rats by kainic acid, picrotoxin, or bicuculline. The
combination of their small size but potent actions makes these toxins a notable class of
compound worthy of in!depth study.
$nother interesting toxin &uite useful in studying receptors for excitatory amino acids in
the mammalian central nervous system is jorotoxin from the venom of Nephila clavata
"3oro spider%. 3orotoxin has been shown to be a specific blocker of the postsynaptic
glutamate receptors, in contrast with bacterial pertuissus toxin that blocks the presynaptic
glutamate receptors. These low molecular weight toxins may provide novel tools for
anticonvulsant research and therapy, demonstrating yet again the potential use of venom
components to be used as investigational ligands or even therapeutics in their own right.
Spider venom peptides
Mu-Agatoxins
The nomenclature of spider peptidic neurotoxins is, unlike that of scorpions, based on the
same convention as that of cone!snails and snakes4 alpha!toxins inhibit the acetylcholine
receptor5 mu!toxins directly abolish muscle action potentials through the inhibition of
muscle sodium channels5 and omega!toxins prevent voltage!activated entry of calcium
into the nerve terminal and the release of acetylcholine.
)pider mu!toxins are cysteine!rich polypeptides, which cause irreversible paralysis and
repetitive action potentials originating in presynaptic axons or nerve terminals. 6linical
effects of a bite from one of the orth $merican funnel web!spider Agelenopsis aperta
are negligible. The venom, however, has yielded several interesting peptides that have
pharmacological properties that may prove to be &uite useful. Two specific mu!agatoxins
have been identified from A. aperta, mu!agatoxin!0 and mu!agatoxin!02, both of which
contain 78 amino acids and four internal disulfide bonds. $lthough these toxins
specifically modify voltage!sensitive sodium channel activity, they have structural
similarities with a plethora of other peptide toxins targeting a myriad of channel types.
$nalysis of these variations may shed light not just on functional residues of the
particular scaffold but also provide data as to the structure and specificity of the binding
sites of the channel being affected. This adds to the emerging wealth of data of a common
scaffold being modified for divergent use.
The omega!agatoxins from A. aperta consist of two subtypes of neuronal calcium
channel toxins with different structural characteristics and binding specificities. Type 0
agatoxins, such as omega!$ga!0, may define a binding site on neuronal calcium channels
that is common to both vertebrates and invertebrates. Type 00 omega!toxins, such omega!
$ga!00, 000 and 02, share limited amino acid se&uence similarity with Type 0 toxins.
9owever, omega!$ga!02$ is able to block omega!6T:!/20$ "Conus geographus%
resistant calcium channels. This use of structurally distinct toxins from common as well
as divergent sources is an excellent manner in which to study a channel.
The peptidic neurotoxins from A. aperta have similar locations of cysteine residues with
neurotoxins from the venom of another orth $merican funnel web spider Hololena
curta as well as neurotoxins from the more distantly related *razilian spider Phoneutria
nigriventer ";andering spider%. This conservation of structure among these three species
is interesting to view from a taxonomic standpoint in that venom peptide se&uences may
be useful as chemotaxonomic tools. Despite being the only lethal species amongst the
three spiders, the toxins of the Phoneutria nigriventer share strong structural similarity,
particularly in the location of the cysteine residues with neurotoxins from these other two
spiders.
;hile the lethal neurotoxin PhTx< from Phoneutria nigriventer venom has a primary
structure shows no homology to any other identified spider toxin, cD$ libraries
constructed from the venom glands revealed that the structure of the preprotoxin initially
synthesized by the Tx< gene shows similarity in se&uence and also in processing with the
synthesis and processing of omega!agatoxin 0$ from A. aperta. Thus, this toxin may not
be &uite so unrelated but rather may represent a case of a spider that is taxonomically
divergent, as P. nigriventer is from the more closely related two . $merican species,
with the venom diverging along with it.
Delta Atracotoxin
=nlike the $merican funnel web spiders, the venoms of $ustralian funnel web spiders are
&uite lethal, consisting of a large number of acute acting neurotoxins. These venoms slow
the inactivation of primate sodium channels causing envenomation symptoms involving
pain at the bite site, salivation, lachrymation, piloerection, generalized skeletal muscle
fasciculation, sweating, nausea, vomiting, diarrhea, pulmonary edema, dyspnoea
followed by respiratory failure, tachycardia and hypertension followed by hypotension
and circulatory failure.
The primary toxic components of the venoms of two species have been isolated and
characterized. The lethal toxins from A. robustus and H. versuta venoms are the >1 amino
acid peptide components robustoxin "atracotoxin% and versutoxin respectively. 2ersutoxin
differs from robustoxin by only ? amino acid residues. Disulfide!bridged cysteine
residues at both the amino! and carboxy!termini and a triplet of cysteines at residues <>!
<8 makes these components unprecedented amongst neurotoxins.

6omparison of rubustoxin and versutoxin.

These presynaptic neurotoxins produce spontaneous, repetitive firing of autonomic and
motor neuron action potentials. 6haracteristic of this autonomic storm is a wave of
endogenous acetylcholine, noradrenaline and adrenaline. These two toxins bind to
tetrodotoxin!sensitive sodium channels, competing with alpha!scorpion toxins. These
toxins cause a voltage dependent slowing of sodium channel inactivation by binding to
the outer surface of the channel, thus interfering with the conformational changes
necessary for gating of the channel.
Latrotoxin
0n contrast to the mostly channel binding small peptide toxins from the $ustralian funnel
web spiders and that from most other spiders, the major bioactive component "latrotoxin%
from the venom of the lethal $merican black widow spider "Latrodectus mactans% is a
much larger protein. ,atrotoxin is responsible for many of the neurological symptoms
clinically seen, causing a massive release of acetylcholine that is stimulated by the
presence of 6a1@ ions. 0t is interesting to compare the activity of latrotoxin with the
smaller peptidic venom component omega!conotoxin. ,atrotoxin is an activator of
synaptosomal calcium uptake, while omega!conotoxin /20$ is an inhibitor of voltage!
sensitive calcium channels of the !type, yet both toxins ultimately produce cramping or
rigid paralysis.