Immunochemical and proteomic technologies as tools for unravelling toxins involved in envenoming by accidental contact with Lonomia obliqua caterpillars.
The accidental contact with Lonomia obliqua caterpillar causes local and systemic symptoms (such as fibrinogen depletion), leading, in some cases, to serious clinical complications (acute renal failure and intracranial haemorrhage). Fortunately, a successful therapeutical approach using anti-Lonomic serum, produced in horses against L. obliqua's bristle extract, has already been put in place. However, a global view of immunogenic toxins involved in the coagulation disorders could help to elucidate the envenoming process. In the present study, our aim was to identify bristle extract's immunogenic components, especially those related to the haemostasis, coupling proteomics and immunochemical approaches (bidimensional electrophoresis, mass spectrometry and immunoblotting). The bidimensional map of bristle extract showed a broad profile of 157 silver-stained spots, where at least 153 spots were immunochemically revealed. Twenty-four of these spots were submitted to sequencing by mass spectrometry and three different categories of proteins were identified: lipocalins, cuticle proteins and serpins. From these protein families, it was observed that the most abundant was the lipocalin family, specifically represented by different isoforms of Lopap (a prothrombin activator protein), reinforcing its relevance during envenoming. Peptide sequences of several other immunochemically revealed spots showed no correspondence to any known sequence and were classified as unknown proteins. These proteins could represent new immunogenic molecules and/or toxins. The sequences presented in this article can be used for oligonucleotide design aiming the amplification of cDNAs coding for new molecules using L. obliqua bristles' cDNA libraries or isolated RNAs as template.
Ricci-Silva
Maria Esther
ME
Laboratory of Biochemistry and Biophysics, Instituto Butantan, Av. Vital Brazil 1500, São Paulo, CEP 05503900, Brazil.
Valente
Richard Hemmi
RH
León
Ileana Rodriguez
IR
Tambourgi
Denise Vilarinho
DV
Ramos
Oscar Henrique Pereira
OH
Perales
Jonas
J
Chudzinski-Tavassi
Ana Marisa
AM
D000595:Amino Acid Sequence; D000818:Animals; D001180:Arthropod Venoms / Q000633:toxicity*; D006487:Hemostasis / Q000187:drug effects* / Q000502:physiology; D015151:Immunoblotting / Q000379:methods; D007120:Immunochemistry / Q000379:methods*; D019476:Insect Proteins / Q000032:analysis / Q000737:chemistry; D007915:Lepidoptera; D054834:Lipocalins / Q000032:analysis / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D009036:Moths; D040901:Proteomics / Q000379:methods*; D012697:Serine Endopeptidases / Q000032:analysis / Q000737:chemistry; D015843:Serpins / Q000032:analysis / Q000737:chemistry
D001180:Arthropod Venoms; D019476:Insect Proteins; D054834:Lipocalins; D015843:Serpins; C047663:cuticle proteins, insects; C119835:Lonomia obliqua prothrombin activator protease; D012697:Serine Endopeptidases