Justin Hessinger

Tess Demeter
 A toxin is a poisonous substance that is
produced by one organism that has negative
effects on another organism

They include small molecules, peptides, or

proteins
 Cause
theyre
freakin
cool!!!
No, but really, we felt that
amatoxins (alpha-amitin) were
interesting because they are the
cause of over 100 human deaths
annually worldwide. The
organisms that are affected by
these seemingly harmless
shrooms suffer mild symptoms
hours after ingestion that
completely disappear only to kill
their unexpecting victims days
later.
 Class of toxins produced by certain types of
Amanita species of mushrooms
Bicyclic octapeptide derivatives
zymogenic until they reach the host

There are at least 10 known variants of

amanitin toxin
Amatoxins account for 95% of fatal
mushroom poisoning
 Alpha-Amanitin is produced by some of the mushrooms
from the genus Amanita.

The most common/deadly species:

Death cap mushroom Destroying

(Amanita phalloides) Angel
(Amanita virosa)
 There are around 600 known species,
some of which are actually edible
They are distributed throughout the world
and found in regions such as;
Mexico
Central Africa
Europe
Canada
South America
United States
 Alpha-amanitin causes inhibition of RNA
Polymerase II by:
strong hydrogen bonding to the bridge helix on
RNA Polymerase referred to as pore 1.
penetrates at pore1 and regulates
translocation and stops elongation at or before
translocation occurs.
it does not affect NTP loading, rather it
prevents the bridge helix from flexing thus
preventing DNA translocation.
 Inhibition of RNA Polymerase II induces stress
activated kinases.
Phosphorylation of p53 protein fraction on serine
residues
This causes suppression of p53 degredation
The accumulated p53 are translocated to
mitochondria and this increases permeability of
the outer membrane which causes the release of
cytochrome C to the cytosol.
This causes programmed cell death.
 Amanitin toxicity is also associated with an
increase in the production of reactive oxygen
species through lipid peroxidation

Increases hepatotoxicity

Although many studies note this reaction

during amanitin toxicity, they are unclear in
stating how amanintin itself induces this
response
 These are all very similar in structure, often
only differing by 1 R-group.

-amanitin Amanullinic Acid

-amanitin Amaninamide
-amanitin Amanin
-amanitin Proamanullin
Amanullin
 Toxicity depends on many factors:
Amount of toxin ingested
Age; multi-organ failure between 6-8 days
ranges
10-20% adults
22-50% children
Immune Function
Exposure Pathway; majority of exposure is
through oral administration, but there are
cases of intravenous injection.
 Although amatoxins are
extremely toxic to humans and
difficult to treat, there are
species that are immune to
these toxins. The mechanisms
of their immunity are highly
misunderstood

Caenorhabditis elegans

Drosophila melanogaster
 Structural mutations of RNAPII
Alpha-amanitin is unable to bind in the proper
location
Most plausible

Metabolic degradation
There are 3 possible modes of degredation
although the most plausible is by cytochrome
P450s

Behavioral Avoidance
 Why are the mushrooms that develop these
amatoxins not affected by them?

Protein precursers are usually used when the protein

is potentially harmful to the host, but needs to be
available on short notice and in large quantities.

The active form of the toxin is not present in the host

organisms.

When entering a target species activation occurs by

cleavage of the 8 terminal amino acids present on the
toxin

Enzymatic reaction catalyzed by prolyloligopeptidase

 The most common pathway is through oral
ingestion

The toxin may be isolated and injected

peritoneally or intravenously, although this
method is really only reserved for lab research
and due to ethicalities, humans are not generally
the test subjects
 For humans:
6-12 hours: incubation (no observable adverse
effects)
Starts with gastrointestinal symptoms
Vomiting, diarrhea, abdominal pain,
hypoglycemia, and dehydration
48-72 hours: evident hepatocellular damage
Coagulopathy and acute liver failure
6-8 days: acute liver and kidney failure
Haemorrhaging, encephalopathy, coma, and
death
 About 10% of individuals exposed to
amanitin in will die in the U.S.

World wide, about 50% of those exposed to

aminitin toxin will die due to inadequate
and/or less accessible medical care

Of those that survive, severe liver and kidney

damage is extremely common and some
patients may die due to complications
pertaining to organ failure
 Silybin
Comes from the Milk Thistle (Silybum marianum)
Competes with amanitin for transport systems in
the plasma membranes in cells
Causes a decrease in penetration of amanitin into
hepatocytes

Thioctic Acid
Powerful antioxidant
Inhibits free radical formation through lipid
peroxidation
Prevents further oxidative tissue damage
 Benzylpenicillin
Inhibits trasporting polypeptide 1B3 in the plasma
membrane which impedes amanitin penetration of
hepatocytes

N-acetylcysteine
Unclear mechanism
May inhibit hepatic uptake of amanitin
May act as strong antioxidant

Cimetidine
Listed as a treatment, but there was no known
mechanism found
 To date it seems that the most effective treatment
involves pumping the stomach right after eating the
toxin.
Because this treatment is extremely time dependant
and symptoms may take hours to show up, its not
largely reliable.

Although there are other proposed treatments, the

ones that we have included are the most common
across notable sources.

The only treatment other than emptying the stomach

content, that has shown any potential benefits
through decrease in hepatic necrosis is Silybin.
These results, as noted in various studies, can be
limited or nullified by the experimental conditions.
There are other studies that have resulted positively
with different toxins, but themselves list a variety of
limitations and problems with their research
 Before eating a random mushroom, there are
a few things you can do:

Meixner Test: tests for presence of amanitin

in the mushroom
Limitations: cannot differentiate between toxic
and hallucinogenic species because psilocin
gives a false positive
Mezlers Reagent: tests for spores of amyloid,
non-amyloid, and dextranoid
Used to identify different types of fungi
 There is potential for therapeutic use against
pancreatic carcinoma
Through conjugation of Amanitin to Anti-
epithelial cell adhesion molecules (EpCAM),
antitumor properties have been shown
EpCAMs are over expressed in many
cancers.
The conjugated molecules are able to bind
these markers and initiate apoptosis
 Although amatoxins are widely distributed
and extremely toxic, they are still very
misunderstood.

The toxic effects suffered from exposure to

amatoxins are difficult to treat and the
mechanisms behind their action is still
unclear. Despite these difficulties, continued
research and understanding is very
important because they may be the key to
future treatments for other illnesses and
survivorship.
 So remember kids,
be careful when
picking your
shrooms!!!

As we understand that peer-reviewing is a time-consuming

process and you may be tired, here is some legal fun you
can have with some awesome shrooms. ENJOY!!!
http://onlinesupermario.com/mario-world.php
Brueckner, F. and Cramer, P. 2008. Structural basis of transcription inhibition by -amanitin and implications for RNA polymerase II translocation. Nature Structural & Molecular Biology. Vol. 15, No. 8: 811-818

Madalan, J., Piotrowska, A., Gomulkiewicz, A., Sozanski, T., Podhorska-Okolow, M., Szelag, A., and Dziegiel, P. 2011. Benzylpenicyllin and acetylcysteine protection from -amanitin-induced apoptosis in
human hepatocyte cultures. Experimental and Toxicologic Pathology. Vol 63: 311-315.

Moldenhauer, G., Salnikiv, A., Luttgau, S., & Harr, I. (2012). Therapeutic potential of amanitin-conjugated anti-epithelial cell adhesion molecule monoclonal antibody against pancreatic carcinoma. JNCI, 1-13.

Nikolova, G., Karamalakova, Y., Hadjibojeva, P., Georgiev, T., Tolekova, A., Gadjeva, V., and Zheleva, A. 2010. Severe mushroom toxin alpha amanitin causes generation of reactive oxygen species in liver
tissues of mice A comparative study by two different instrumental methods. Trakia Journal of Sciences. Vol. 8, No. 2: 149-154.

Springer, M. 2006. Effect of Alpha-Amanitin. Retrieved April 16, 2012 from <http://www.chem.uwec.edu/Webpapers2006/sites/springmg/amanitin.html>

Stump, A., Jabonski, S., Bouton, L., and Wilder, J. 2011. Distribution and mechanism of -amanitin tolerance in mycophagous Drosophila. Environmental Entomology. Vol. 40, No. 6: 1604-1612.

Thiel, C., Thiel, K., Klingert, W., Diewold, A., Scheuermann, K., Hawerkamp, E., Lauber, J., Scheppach, J., Morgalla, M., Konigsrainer, A., and Schenk, M. 2011. The enterohepatic circulation of amanitin:
Kinetics and therapeutical implications. Toxicology Letters. Vol. 203: 142-146.

Tsao, D., Park, N., Nag, A., and Martinson, H. 2012. Prolonged -amanitin treatment of cells for studying mutated polymerases causes degradation of DSIF160 and other proteins. RNA Society. Vol. 2: 222-
229

Tong, Tri C. "Comparative Treatment of ."Annals of Emergency Medicine. 50. (2007): 282-288. Print.

Zheleva, A., Gadjeva, V., and Zhelev. M. 2003. Free radical formation might contribute to the severe amatoxin hepatotoxicity. Trakia Journal of Sciences. Vol. 1, No. 3: 42-45.

mushroom poisoning syndromes. (2012). Retrieved from http://www.namyco.org/toxicology/poison_syndromes.html