Estudo comparativo da fração de baixa massa molecular de venenos viperídeos do Brasil e de outros locais

Abstract

Snakebites were classified by WHO as neglected diseases, and the study of venom content from snakes that cause accidents is very important and has a great impact on public health. Venomics comprises a set of analytical techniques that allows the study of the toxins present in the venom, understanding their structures and their biological functions and evolutionary relationships. However, when it comes to low mass molecules little is known about this fraction due to the lack of effective approaches for their study. Thus, the present study aimed to apply different methods of preparation and separation for the study of low mass molecules (<10kDa), to suggest alternatives for a better characterization of this little explored fraction. Viperidae of different species/genus and biomes were processed by ultrafiltration and subjected to hydrophilic interaction liquid chromatography (HILIC), mass spectrometry and antimicrobial activity screening. This is the first work to use the HILIC method for low molecular mass characterization of snake venoms, which showed better separation and molecular diversity than the classical RP-HPLC method. The fractions showed similar chromatographic profiles for snakes of the same genus, but genus-specific features were also detected. Mass spectrometry analyses revealed the presence of canonical BPPs, new peptides, and protein fragments (possible cryptides), after HILIC fractionation. the low molecular fractions from Agkistrodon bilineatus, Bitis gabonica, Bitis arietans and Bitis nasicornis showed antimicrobial activities against bacteria and fungi. In summary, the present study shows that HILIC is an efficient approach for the biochemical characterization of low mass molecules showing relevant biological activities from snake venoms, that could also be a new source of candidate-molecules with pharmacological potential.