Fungal Toxins

NST110, Toxicology
Department of Nutritional Sciences and Toxicology
University of California, Berkeley

Outline
1. Aflatoxin
2. Ergotism
3. Psilocybin
4. -amanitin
5. Phytoalexins
6. Muscarine

Aflatoxin
Aflatoxinfrom Aspergillus flavusactive ingredient is Aflatoxin B1.
Symptoms: hepatotoxin at high doses, carcinogenic at low doses (most potent liver
carcinogen).
FDA allowable amount is 15 ppb in food products and 100 ppb in feed.
Mechanism of actionaflatoxin is a potent mutagen

Toxicity and Detoxification of Aflatoxin

Kensler T W et al. Toxicol. Sci. 2011;120:S28-S48

Biotransformation pathways for aflatoxin B1.

Aflatoxin and Hepatitis

Relative risk
AFB1 metabolites in urine
Hep B virus antigens
AFB1 + hepB
N7 guanine adduct + HepB

3.4
7.3
60
>120

Aflatoxin is a mutagen that initiates cells and can lead to liver cancer.
Hepatitis B causes chronic liver inflammation which can promote cells through ROS and
can also lead to liver cancer.
Aflatoxin (initiation) + hepatitis B (promotion) can lead to further liver cancers.

Key steps in the development of HCC by viral (HBV and HCV) and chemical (aflatoxin) factors.

Kensler T W et al. Toxicol. Sci. 2011;120:S28-S48

The Author 2010. Published by Oxford University Press on behalf of the Society of Toxicology.
All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Potent Protection against Aflatoxin-Induced Tumorigenesis through

Induction of Nrf2-Regulated Pathways by the Triterpenoid 1-[2-Cyano3-,12-Dioxooleana-1,9(11)-Dien-28-Oyl]Imidazole

Triterpenoid 1-[2-Cyano-3-,12Dioxooleana-1,9(11)-Dien-28Oyl]Imidazole; CDDO-lm

Aflatoxin

* CDDO-lm

Cancer Res. 2006 Feb 15;66(4):2488-94

Protection by CDDO-Im against aflatoxin-DNA adduct

formation

Protection by CDDO-Im against AFB1-induced GST-P

positive foci formation
GST-P: putative preneoplastic lesions

Nrf2: control of sensitivity to carcinogens

Ahr/Arnt

Ahr/Arnt

Ergotism
Ergotfrom Claviceps purpureagrows as sclerotia on individual rye kernels .
Symptoms: sometimes one or both
gangrenous typeblood flow stops, mummified extremities and loss of limbs, painful.
Convulsive typeneurologicalnumbness, blindness, convulsions, paralysis.

Active substances
ergotamine (LD50 10 ug/kg rodents):
LSD has ethyl groups on the R (lysergic acid).

O
R 2N

C H3

N H

Ergotamine/Dihydroergotamine

-a class of medications called ergot alkaloids

-used to treat migraine
-constrict intracranial blood vessels (vasoconstriction)
reduce neruogenic inflammation (a partial agonist )
5-HT1D receptor and alpha-adrenergic receptor
-also acts as a partial agonist for central 5HT1A receptors (nausea and
dysphoria), dopamine D2 receptors (nausea and vomiting)

Claviceps gained great notoriety when Albert Hoffman (Sandoz Labs, Switzerland) in 1938
discovered that the sclerotia also contained lysergic acid amide, a precursor of LSD (Lysergic
acid diethylamide), one of the most potent psychotropic drugs. He was the first to synthesize
and try LSD!

* Feelings of "strangeness"
* Vivid colors
* Hallucinations
* Confusion, panic, psychosis, anxiety
* Emotional reactions like fear, happiness or sadness
* Distortion of the senses and of time and space
* "Flashback" reactions * Increases in heart rate and blood pressure
* Chills
* Muscle weakness
LSD is Schedule I in the United States. This means it is illegal to manufacture, buy, possess,
process or distribute LSD without a DEA license.

LSD is agonist of many subtypes of 5-HT receptor, and possibly dopamine receptors.

5-Hydroxytryptamine (Serotonin) Receptor

1. Affect various neurological processes: aggression, anxiety, appetite,
cognition, learning, memory, mood, sleep, nausea, thermoregulation.
2. Modulate the release of neurotransmitters and hormones: glutamate,
GABA, acetylcholine, dopamine, norepinephrine/epinephrine, cortisol,
substance P, oxytocin, vasopressin, etc.
3. Serotonin receptors are targets of pharmaceutical and illicit drugs: antidepression, anti-migraine, anti-emetics, anti-psychotics, hallucinogens,
entactogens (chemicals that induce the feelings of empathy). Nearly all
known 5HT2A agonists produce hallucinogenic effects in humans.

5-Hydroxytryptamine (Serotonin) Receptor

Receptor
5-HT1

Action
Gi/Go; decrease intracellular cAMP
Gq/G11; increase intracellular IP3 and Diacylglycerol

5-HT3

Ligand-gated sodium and potassium channel

5-HT4

Gs; increase intracellular cAMP

5-HT5

Gi/Go; decrease intracellular cAMP

5-HT6

Gs; increase intracellular cAMP

5-HT7

Gs; increase intracellular cAMP

LSD is an agonist of 5-HT1A, 5-HT2A, 5-HT2C, 5-HT5A, 5-HT5, 5-HT6.

Signal Transduction Pathway of 5HT2 Receptor

PLC: phospholipase C, PIP2: Phosphatidylinositol 4,5-bisphosphate

IP3: inositol 1,4,5-trisphosphate, DAG: Diacylglycerol, PKC: protein kinase C

Psilocybe cubensis
is a species of psychedelic mushroom whose principle active compounds are
psilocybin and psilocin.
Mode of Action: Psilocybin is rapidly dephosphorylated in the body to psilocin which
then acts as an agonist at the 5-HT2A serotonin receptor in the brain where it mimics
the effects of serotonin (5-HT). About 1% of the weight of Psilocybe cubensis
mushrooms is Psilocybin.
psilocybin

Psilocybin-containing mushrooms are commonly called magic mushrooms or simply "shrooms".It is

isolated by Albert Hoffman. Effects of psilocybin are comparable to those of a shorter LSD trip.

Psilocybin binds to 5-HT2A with high affinity, and also 5-HT1A, 1D, 2C with lower
affinity. It may also act through dopamine receptors.
LD50 is 280 mg/kg BW, about 1.5 times that of caffeine.

L-Tryptophan is the precursor for biosynthesis of serotnin in vivo and

for synthesizing psilocybin in vitro

Death Cap Mushroom

Amanita phalloidesDeath cap
Abdominal pains, diarrhea, vomiting (1-2 days)
Liver failure; can cause death
Treatment: liver transplant or plasmaphoresis
Toxic agents: amanitinsspecific RNA polymerase II inhibitorGI tract is where
first symptoms occur, followed by liver destruction.
-amanitin is a cyclic nonribosomal peptide of eight amino acids.

Mechanism of alpha-amanitin Action

The amanitin binding site is

beneath a "bridge helix"
extending across the cleft
between the two largest pol II
subunits, Rpb1 and Rpb2, in a
"funnel"-shaped cavity in the
pol II structure.

From the crystal structure solved, -amanitin interacts with the bridge helix in
the RNA polymerase II (pol II). This interaction interferes with the translocation
of RNA and DNA needed to empty the site for the next round of RNA synthesis.
The addition of -amanitin can reduce the rate of pol II translocating on DNA
from several thousand to a few nucleotides per minute, but has little
effect on the affinity of pol II for nucleoside triphosphate, and a
phosphodiester bond can still be formed. Bridge helix is designed to be
flexibile for its movement is required for translocation. Binding of -amanitin
puts a constraint on its mobility, hence slowing down the synthesis of RNA.
Bushnell, David A. et al. (2002) Proc. Natl. Acad. Sci. USA 99, 12181222

Mechanism of alpha-amanitin Action

There is a strong hydrogen
bond between hydroxyproline 2
of alpha -amanitin and bridge
helix residue Glu-A822.

The derivative proamanullin,

which lacks the hydroxyl group
of hydroxyproline 2, involved
in hydrogen bonding to bridge
helix residue Glu-A822, is much
less effective than alpha -amanitin

Phytoalexinschemical produced in defense of attack from another plant or organism,

or even physical stress or damage.
1. Potatoesif potatoes are infected with a fungus, the fungus eventually stops
growing.
-Active component is solanine (AChE inhibitor)
-Solanine has both fungicidal and pesticidal properties, and it is one of the plant's
natural defenses.
-Solanine poisoning is primarily displayed by gastrointestinal and neurological
disorders. Symptoms include nausea, diarrhea, vomiting, stomach cramps, burning of
the throat, headaches and dizziness. Hallucinations, loss of sensation, and paralysis,
fever, jaundice, dilated pupils and hypothermia have been reported in more severe
cases.

2. Sweet potatoesIpomea batatasmoldy sweet potato causes liver poisoning due

to a toxic agent produced by sweet potatodark ring inside sweet potato indicates
there was fungal growth
Active component: ipomeamarone, ipomeanol

Ipomeanol inhibits CYP3A4