BACTERIAL SERPINS: PRODUCTION, CHARACTERIZATION AND ACTIVITY AGAINST MELANOMA CELLS

Abstract

Serpin is the name given to the superfamily of proteins with a wide variety of biological functions, and that has as main characteristic the inhibition of serine proteases. Our group recently described and characterized the first of the serpin Sphenophorus levis, popularly known as "bicudo de cana", which attacks the Brazilian sugar cane causing great damage to this crop. Until recently it was believed that the serpins were unique to eukaryotic organisms. However, with the development of techniques for sequencing of genomes, these proteins also came to be identified in prokaryotic organisms. Thus, this project proposes the cloning, expression and biochemical characterization of the inhibitory activity of bacterins serpins six described in the database of the genome "genbank". The serpins that will be the subject of our study were selected according to the amino acid sequence of its RCL (reactive center loop), to produce serpins with different specificities for a wider range of proteases. In addition, we also propose to produce recombinant serpins with site of mutations directed at amino acid P1 of the RCL, which is an amino acid important for the specificity of these proteins against different proteases. To complete the project, also intend to study the effect of these six serpins in the proliferation of melanoma cells, it was reported that some serine proteases of the tissue kallikrein family of operating in the development of this cell type.Thus, we can also say that this project has the objective of obtaining recombinant serpins specific for certain serine proteases, such as the humans tissues kallikreins 1, 3, 5, 6 and 7, that will assist us in studies to better understand the role physiological and/or pathological, these proteases in different tissues where they appear expressed, and can serve as candidates for new therapeutic procedures.