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23 Congress of the International Union for Biochemistry and Molecular Biology
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44 Annual Meeting of the Brazilian Society for Biochemistry and Molecular Biology
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th
Foz do Iguaçu, PR, Brazil, August 24 to 28 , 2015
VENOM PROTEOMICS AND TRANSCRIPTOMICS FROM BRAZILIAN SPIDERS
Thiago Ferreira de Abreu1, Rafael Lucena Lomazi1, Milton Yutaka Nishiyama
Junior2, Ursula Castro de Oliveira2, Josias Falararo Pagotto1, Eduardo Shigueo
Kitano2, André Zelanis3, Solange M. T. Serrano2, Inácio Junqueira de Azevedo2,
Pedro Ismael da Silva Jr2, Alexandre Keiji Tashima1
1. Departamento de Bioquímica, Escola Paulista de Medicina, Universidade Federal de
São Paulo, São Paulo, Brasil; 2. Centro de Toxinas, Resposta-Imune e Sinalização
Celular (CeTICS), Instituto Butantan, São Paulo, Brasil; 3. Instituto de Ciência e
Tecnologia, Universidade Federal de São Paulo, São José dos Campos, Brasil.
Introduction and Objectives
The class Arachnida is amongst the most diverse terrestrial groups of organisms. It is
also one of the most successful groups of venomous animals on the planet. One factor
contributing to their success is the production of a toxic secretion , which is used for
subduing prey as well as for protection against predators. Considering the biological
potential of the molecules in the venoms of Brazilian spiders, we conducted a proteomic
and transcriptomic study of Acanthoscurria juruenicola and Nephilengys cruentata and
unveiled new proteins, enzymes and peptides with potential antimicrobial activity.
Materials and methods
Venoms were extracted by electrical stimulation and stored at -80°C until use. Proteins
were digested with trypsin at 37°C overnight or with thermolysin at 75°C for 45 min.
Peptides were separated in a capillary C18 column, ionized by electrospray and
analyzed by data dependent and data independent LC-MS/MS. Proteins were identified
by search against predicted proteins database derived from the Trinity-assembled
transcripts made from venom glands. cDNA libraries were sequenced on an Ilumina
HiSeq1500 sequencer.
Results and conclusions
Combined proteomic and transcriptomic analyses resulted in the identification of 161
and 353 proteins in the venoms of N. cruentata and A. juruenicola, respectively.
Sequences were homologous to cysteine-rich venom proteases, metalloproteases,
carbonic anhydrases, hyaluronidases, chitinases and theraphotoxins. A. juruenicola
toxins showed high similarity to proteins identified in other spiders of the Acanthoscurria
genus, but they also resembled toxins from Grammostola spiders. Absolute
quantification indicated that chitinases and metalloproteases were the most abundant
proteins in the venom of N. cruentata. Moreover, preliminary results indicate that a new
theraphotoxin from A. juruenicola possesses antimicrobial activity. Other components in
the mass range of 3-8 kDa have also been found in the venoms and experiments are in
progress for their characterization.
Acknowledgements
FAPESP and CAPES
Key Words (only three words)
Proteomics, transcriptomics, spider
Brazilian Society for Biochemistry and
Molecular Biology (SBBq)