Venomics, what else?

Eager to unlock the molecular secrets of venoms?Venoms, Venomics and Antivenomics

Venomic and antivenomic tools for exploring snake venom proteomes and improving the management of snakebite envenoming, a neglected pathology

Laboratorio de Venómica y Proteinómica Estructural

Gema Bolás Jordi Durban

Libia Sanz Alicia Pérez

Juanjo Calvete

Carolina Llorente

Beatriz Companys

Raquel SanzDavinia Plá

Carlos Corrêa-NettoIVB, Niterói, RJ

Johara Boldrini-FrançaUniversidade de São Paulo

Ribeirão PretoRenata S. RodriguesUniversidade Federal de Uberlândia

Marliete M.S. SilvaUniversidade Federal Rural de Pernambuco

ChemaBruno

Yamileth

Rob Harrison

David Theakston

Proteomics and antivenomics of Echis, Bitis, and Naja venoms

David Warrell

S. c. cat enat us

S. c. t ergemi nus

S. c. edwardsi i

S. c. cat enat us

Stephen P. Mackessy

H. Lisle Gibbs

And just like that, Larry Lizard found he`d evolved into a snake!

100 Mya...

Boideae

Advancedsnakes

Viperidae

Elapidae

Atractaspididae

Colubrinae

Venom represented a keyinnovation in ophidian evolutionthat allowed advanced snakes totransition from a mechanical(constriction) to a chemical(venom) means of subduing anddigesting prey larger thanthemselves.

Evolution of venom-delivery systems

Early evolution of the venom system in lizards and snakesFry, BG et al. Nature 439, 584-588 (2 February 2006)

2x

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Gene multiplication

Multigene protein family

Natural Selection

- Accelerated evolution- Neofunctionalization

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... A C V G G H Y F V C D S E R C N ...

... A C V F G H Y F V C D S E R C N ...

... A C V G G N Y F A C D S E R C N ...

Accelerated mutation rate

Protein sequence diversity- Modulation of function- Adaptive role

Low abundance proteins may be moreplastic. A subset may serve to“customize” an individual venom tofeeding on particular prey or mayrepresent orphan molecules evolvingunder neutral selection "in search for afunction”

Abundant venom proteinsmay perform generic killingand digestive functions.

Snake venomics

- Reduce universe of toxin scaffolds- Extensive immune crossreactivity Offers the possibility of defining the

minimal set of venoms containingthe epitopes necessary to generatetherapeutic broad-range polyvalentantisera.

Venom phenotyping

Window of opportunity to expandthe range of clinical use of currentantivenoms

Antivenomics

Nothing in Biology Makes Sense Except in the Light of EvolutionThe American Biology Teacher, 35:125-129 (1975)Theodosius Dobzhansky (1900-1975)

The knowledge of evolutionary trends along with venom phenotyping may be used to replace the traditionally used phylogenetic hypothesis for antivenom production strategies by cladistic clustering of venoms based on proteome phenotype and immunological profile similarities.

ColombiaEcuador

Costa Rica

Venezuela10-15 Mya

Rapid dispersal and diversification of Bothropsacross South America may have occurred throughadaptive radiations into habitats devoid of viperid

competitors

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B. asper juvenile

B. asper adult

Potency vs. Plasticity

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Addressing evolutionary trends...

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Cmm

Elda E. Sánchez, John C. PérezNatural Toxins, Texas

Type I venoms: SVMP; toxicity(ontogenetic)Type II venoms: toxicity; SVMP(paedomorphic)

g g _ _ C18 for veneno2 B nigroviridis 2mg 110909:1_Conc C18 for veneno2 B nigroviridis 2mg 110909:1_Inject

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J. Proteome Res. 7, 2445-2457 (2008) J. Proteome Res. 9, 4234-4241 (2010)

Preinmune serum

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Alicia Pérez

S I II II

Trivalent antiserum-4

Clementino de Lima, D. et al. (2005)eCAM 2(1) 39-47

Mechanism of action of ACE inhibitors developed from a bradykinin-potentiating enzymeisolated from the venom of the Brazilian pitviper (Bothrops jararaca) and approved in 1979by the FDA to treat high blood pressure and heart disease. Angiotensin-I causesvasoconstriction which raises the blood pressure.

Cushman DW, Cheung HS, Sabo EF, Rubin B, et al. 1979.Development of specific inhibitors of angiotensin I convertingenzyme (kininase II). Fed. Proc. 38: 2778–82.

CaptoprilSergio Henrique Ferreira

Ferreira, S. H. A Bradykinin-potentiationg factor (BPF) present in the venom of Bothrops jararaca.

British Journal of Pharmacology, v. 24, p. 163-169, 1965.

Cone Snail Venom Effective Remedy For Pain

The synthetic form of the Conus magus peptide toxin, ω-conotoxin MVIIA, ziconotid (Prialt®), marketed byElan Pharmaceuticals, was approved by the FDA in 2004 to be administered as an intrathecal infusion totreat severe chronic pain in patients who are intolerant or refractory to other systemic analgesics such asmorphine. Ziconotide blocks voltage gated N-type calcium channels in the spinal cord and reduces therelease of pronociceptive neurotransmitters in the dorsal horn of the spinal cord, resulting in an inhibitionof pain signal transmission

McIntosh M, Cruz LJ, Hunkapiller MW, Gray WR, OliveraBM (1982). Isolation and structure of a peptide toxin fromthe marine snail Conus magus. Arch. Biochem. Biophys.218:329-34.

The work leading to Exenatide started with the detection and isolation of exendin-3 fromthe venom of the Mexican bearded lizard (Heloderma horridum) by John Eng andcolleagues (Eng et al., 1990). Exendin-3 is a pancreatic secretagogue of the glucagonsuperfamily. Exendins can bind and activate the human glucagon-like peptide 1 (GLP-1)receptor to enhance glucose-dependent insulin secretion, suppressing elevated glucagonsecretion. Thus, can be used in the management of Type 2 diabetes mellitus. The FDAapproved the use of exenatide in 2005.

Eng J, Kleinman WA, Singh L, Singh G, Raufman JP (1992).Isolation and characterization of exendin-4, an exendin-3analogue, from Heloderma suspectum venom. Furtherevidence for an exendin receptor on dispersed acini fromguinea pig pancreas. J. Biol. Chem. 267:7402-5.

Anti-angiogenesis: a new concept for therapy of solid tumorsFolkman J.Ann Surg. 1972 Mar;175(3):409-416

99mTC-Bitistatin

125I-Bitistatin

5 µg

5 µg

Effect on the adhesion of α1-K562 to coll IV () and α2-K562 to coll I (o).

IC50 = 0.8 nM

AI, Angiogenesis index = nº vessel branch points

60 ± 9 79 ± 11

179 ± 23

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84% Inhibition

n = 4

Effect of obtustatin () and PBS () on Lewis lung carcinoma growth in C57BL/ 6 mice

Cezary Marcinkiewicz

Conditioned Transgenic mice for jerdostatin

Gema Bolás

Rophalorus junceus

La forma más habitual de preparación de una dilución homeopática es la siguiente: se coge 1 ml de la sustancia original yse mezcla con 99 mL de agua. Se agita este preparado y se obtiene una dilución de 1 CH (Centesimal de Hahnemann).Elnúmero representa la concentración del medicamento. Cuanto más alto es, más diluído está, siendo el número lacantidad de ceros: Por ejemplo, 4CH quiere decir que está diluído 1:10000, 5CH, sería 1:100000, y así sucesivamente.30CH = 1:1060

Asumiendo 100 mg toxinas (= 6 x 1018 moléculas) /ml veneno, 9CH equivale a 0.6 moléculas por ml; 10CH = 0.2moléculas/frasco; Se necesitarían 10101 frascos para encontrar una sóla molécula de toxina de Rophalorus junceus...¡VidatoX es AGUA!

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