IIB   20738
INSTITUTO DE INVESTIGACIONES BIOLOGICAS
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Cloning, expression and inhibitory activity on neuroblastome cell growth of Helja, the jacalin-like lectin from sunflower
Autor/es:
GENOULA M ; DE LA CANAL L; SILVEYRA X; CARVALHO A; REGENTE M; DEL RIO M; GOMES V; PINEDO M
Lugar:
Foz do Iguazú
Reunión:
Congreso; 11 Congreso Internacional de Biología Molecular de Plantas; 2015
Resumen:
The lectins are a group of proteins that specifically bind different sugar motifs. They are classified in 12 families according to their sequence and structural similitude. In plant, their endogenous function has been related to biotic and abiotic stress responses. A range of biomedical applications that include antiviral and antifungal activities as well as the development of kits for cancer diagnosis has also been reported for these proteins. Helja, a sunflower jacalin-like lectin able to bind mannose, was purified in our laboratory and characterized regarding its anti-Candidiasis capability. To further assess its putative biotechnological applications, Helja was cloned, expressed and analyzed in its ability to recognize neuroblastome cells.The Helja partial sequence obtained through a proteomic analysis altogether with data mining let us to identificate a scaffold putatively encoding for Helja. This sequence was used to design primers to amplify, by reverse transcription PCR (RT-PCR), a complete coding sequence (CDS) with 5´and 3´ends compatible with the cloning in the expression vector pET-32Ek/LIC. Thus, transformed cells induced with IPTG expressed a protein of 35 kDa, consistent with the MW of the chimera named rHeljaT. The identity of rHeljaT was verified by protein blot using specific antibodies before the purification through Ni and mannose affinity chromatography. Digestion with entorokinase released the jacalin-like portion (rHelja), which in turn, inhibited the growth of neuroblastome cells and disturbed the morphology and the extracellular matrix of these cells. Therefore, rHelja should be further studied as a tool to control the development of neuroblastome tumors.