CHARACTERIZATION OF THE LACHESIS MUTA VENOM BY A PROTEOMIC APPROACH

FUKUSHIMA, JORGE; BISCOGLIO, MIRTHA; RIVERA, ELENA.

Valencia, España

Congreso; Joint Congress SEProt-EuPA on Proteomics & Pathology -from both sides of the; 2007

SEProt-EuPA

Lachesis muta venom causes - at a very low concentration (4 ng/ml) – several biological responses with potential clinical applications such as the ability of modulating the activity of antioxidant enzymes and the expression of genes related to angiogenesis and apoptosis. “In vivo” brings about up-regulation of innate and T-mediated immunity and interferon gamma release, factors leading to the inhibition of the growth of tumors.We undertook the study of venom proteomics. 2D SDS-PAGE and RP-HPLC-MS of total venom were performed to study its complexity degree. Results show the presence of proteins ranging between 14 and 32 kDa as well as low molecular mass components (1-1.4 kDa). The venom was then chromatographed through a Superdex G75 column; all fractions obtained were purified by RP-HPLC and peaks submitted to ESI-MS. Fractions showing the presence of only one or few molecular species (these submitted to further purification) were subjected to             N-terminal Edman degradation and sequence alignments studies were carried out. Several A2 phospholipases were detected. Different biological activities of the purified fractions are being performed for the purpose of defining venom components involved in the various  responses observed, all this as a contribution to the development of forthcoming clinical applications.                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           Lachesis muta venom causes - at a very low concentration (4 ng/ml) – several biological responses with potential clinical applications such as the ability of modulating the activity of antioxidant enzymes and the expression of genes related to angiogenesis and apoptosis. “In vivo” brings about up-regulation of innate and T-mediated immunity and interferon gamma release, factors leading to the inhibition of the growth of tumors.We undertook the study of venom proteomics. 2D SDS-PAGE and RP-HPLC-MS of total venom were performed to study its complexity degree. Results show the presence of proteins ranging between 14 and 32 kDa as well as low molecular mass components (1-1.4 kDa). The venom was then chromatographed through a Superdex G75 column; all fractions obtained were purified by RP-HPLC and peaks submitted to ESI-MS. Fractions showing the presence of only one or few molecular species (these submitted to further purification) were subjected to             N-terminal Edman degradation and sequence alignments studies were carried out. Several A2 phospholipases were detected. Different biological activities of the purified fractions are being performed for the purpose of defining venom components involved in the various  responses observed, all this as a contribution to the development of forthcoming clinical applications.