Evolutionary Convergence of Exotoxins

Fatima Sheikh
Outline

• Keywords 
• Exotoxins
• Types of Exotoxins
• Diphtheria
• Botulinum
• Tetanus From http://forgetfulvetstudent.blogspot.ca/2015/09/exotoxin-vs-endotoxin.html

• Take Home Message

Why are exotoxins important?

• Many bacteria produce toxins, but not all affect humans (e.g. BT
toxin targets some insects harmful to crops)
• Endotoxins vs. exotoxins
• Gram positive and gram negative battery produce exotoxins
• Named based on: target cell, species of bacteria or disease,
function, assigned letter, or type of toxin
• Convergent evolution: many exotoxins follow same basic
mechanism
Types of Exotoxins

• Type I (superantigens)
• Superantigen binds to MHC class II cell surface
and T-cell receptors
• Elevated levels of cytokines (e.g. IL-2)
• Not translocated (i.e. do not enter cell)

Fig. 9-6

From Salyers & Whitt, 2002

Types of Exotoxins

• Type II (membrane-disrupting toxins)

• Lyses host cell, disrupting structural integrity of membrane
• Two types:
• Forms protein channel in membrane of host cell
• Osmosis used to rupture cell
• Enzyme that destabilizes phospholipid bilayer
• E.g. some phospholipases remove charged phosphate head group from phospholipids
Fig. 9-5a

From Salyers & Whitt, 2002

Fig. 9-5b

From Salyers & Whitt, 2002

Types of Exotoxins

• Type III (A-B toxins)

• Contains to components: binding portion (B) and enzymatic portion (A)
connected by disulfide bonds
• Toxic portion remains inactive until disulfide bonds are cleaved
• Two types: 
• Simple
• Single polypeptide
• Complex
• B portion is composed of multiple subunits, A is separate polypeptide
 Fig. 9-3

From Salyers & Whitt, 2002

Fig. 9-4

From Salyers & Whitt, 2002

Modification of regulatory
host GTPase

• GTPases belong to the RHO

family which plays an
important regulatory role in
signalling pathways of
necessary functions
• Development, activation,
and function of B and T-
cells 
• Migration and phagocytosis
• Sensing chemotactic
gradients
• Degranulation and NADPH
oxidase activity
From Aktories, 2011
ADP-ribosylation of RHO
From Aktories, 2011
GTP-bound RHOA

From Aktories, 2011

From Aktories, 2011
 https://en.wikipedia.org/wiki/Corynebacterium_diphtheriae

Diptheria toxin

• Produced by Corynebacterium diphtheriae 

• Gram positive, non-spore forming, nonmotile,
aerobic rod
• Bacteria acquired by inhalation
• Colonizes in throat
• Grayish pseudomembrane (fibrin, bacteria,
inflammatory cells) forms in throat that adheres to
tissue; causes underlying tissue to bleed when removal
attempted
• Irregular heartbeat, difficulty swallowing, damage to
interior organs, stupor, coma, eventually death
Diphtheria

(From Sandvig & Deurs, 2005)

 https://www2.le.ac.uk/project
s/vgec/highereducation/topics
/microbial-genetics-
1/infection/toxinst

Fig. 9-10

From Salyers & Whitt, 2002

Botulinum toxin

• Produced by Clostridium botulinum 

• Gram-positive, spore-forming, obligate
anaerobe, rod
• Acquired by ingestion
• Attacks peripheral neurons and
blocks neurotransmitter release
• Nausea, vomiting, headache, double
vision, slurred speech, neurological
symptoms, general flaccid paralysis
https://microbewiki.kenyon.edu/index.php/Clostridium_botu
linum_Neurotoxins
 https://openi.nlm.nih.gov/detailedres
ult.php?img=PMC3206617_toxins-02-
00024-g001&req=4

Botulinum
Tetanus toxin

• Produced by Clostridium tetani

• Gram-positive,spore-forming, motile,
obligate anaerobe, rod
• Attacks central nervous system and
inhibits neurotransmitter release
• Causes spastic paralysis

https://microbewiki.kenyon.edu/index.php/Clostridium_tetani
Tetanus

From Turton et al, 2002

Take Home Message

• 3 types of toxins, but type III (A-B toxins) are most common
• Same general mechanism, but different results (botulism causes paralysis while
tetanus causes convulsions)
• Differences are result of variations in which receptor B protion binds to, how A
portion enters host cell (membrane translocation), and which processes portion A
affects
• B portion of Diphtheria binds to HB-EGF, endocytosis allows A portion to
translocate into host, which catalyzes ADP-ribosylation of EF-2
• Botulin toxin and tetanus are zinc-dependent metalloproteases that
inhibit release of neurotransmitters, but the former results in flaccid
paralysis, the latter in spastic paralysis
References

• Aktories, K. (2011). Bacterial protein toxins that modify host regulatory GTPases. Nature

Reviews Microbiology, 9, 487-498. 
• Rossetto, O., Pirazzini, M., Bolognese, P., Rigoni, M., Montecucco, C. (2011). An update on the
mechanism of action of tetanus and botulinum neurotoxins. Acta Chim. Slov., 58, 702-707.
• Rossetto, O., Pirazzini, M., & Montecucco, C. (2014). Botulinum neurotoxins: genetic,
structural and mechanistic insights. Nature Reviews Microbiology, 12, 535-449.
• Salyers, A. A., & Whitt, D. D. (2002), Bacterial pathogenesis: A molecular approach.
Washington, DC: ASM Press.
• Sandvig, K., & Deurs, B. v. (2005). Delivery into cells: lessons learned from plant and bacterial
toxins. Gene Therapy, 12, 865-872.
• Turton, K., Chaddock, J. A., & Acharya, K. R. (2002). Botulinum and tetanus neurotoxins:
structure, function and therapeutic utility. Cell, 12(11), 552-558.