Study of protein expression in cells treated with an antiapoptotic protein, derived from Lonomia oblique hemolymph

• Grant number: 09/06511-1
• Support Opportunities: Scholarships in Brazil - Master
• Start date: October 01, 2009
• End date: September 30, 2011
• Field of knowledge: Biological Sciences - Biology
• Principal Investigator: Ronaldo Zucatelli Mendonca
• Grantee: Luciana Moreira Martins
• Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil

Abstract

Study of protein expression in cells treated with an antiapoptotic protein, derived from Lonomia oblique hemolymph.The system of proteins expression using Baculovirus vector has been intensively used for the production of recombinant proteins. This system has several advantages, including the production of recombinant proteins with functional immunogenic active, allowing the formation of post-translacionais changes and to contain one strong promoter (poliedrin). It has been reported that this system allows the production of high levels of recombinant proteins, which reach up to 50% of total cell protein. However, in some cases, this system of expression produces less than 2 orders of magnitude of protein secreted when using Sf-9 cells. The infection of cells by baculovirus induces death by apoptosis, which limits the production of recombinant proteins. Thus, one way to increase productivity or reduce cell would inhibit this cell death. Recently the staff of our laboratory reported the occurrence of cell death by apoptosis in cultures of mammals and insects cells in depleted nutrients cultures or induced by chemical agents. They have showed that supplementation of insect or mammalian cell cultures with hemolymph of Lonomia obliqua may extend the viability of cell culture to avoid the death by apoptosis. In this work was isolated and purified the protein responsible for this effect. The addition of this protein in the culture, in death process by apoptosis, allowed the extension of cellular life leading to greater viral replication and hence a greater production of recombinant proteins expressed by baculovirus. For the broad spectrum of activity of our protein, as in insect cells, as in mammals, either by induction of apoptosis by chemical agents, viral or nutrition, we believe that this protein may act as a suppressor of the apoptosis process, by a via conserved in different organisms. If so, this protein could possibly act in synergism with the P35 in the baculovirus or other inhibitor of apoptosis cytological, permitting greater viral replication and expression of recombinant proteins, with an action delayed time in which the action of P35 is not able to prevent cell death. The precise knowledge of proteins involved in anti apoptotic process can be very important in defining the mechanisms of action of the protein under study. Therefore, the objective of this work is to do an analysis of the expression of proteins in cells, treated or not with the anti apoptotic protein, obtained from hemolymph of Lonomia obliqua after induction or not with chemical or viral inducers of apoptosis.

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

MARTINS, Luciana Moreira. Effect of an anti-apoptotic protein obtained from the hemolymph of Lonomia obliqua on the mitochondria of cells Sf-9.. 2011. Master's Dissertation - Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI) São Paulo.