Maisa Splendore Della Casa

Publications (2)4.68 Total impact

• Article: Engineered mammalian vector to express EGFP-tagged proteins as biomarkers.

  Geraldo Santana Magalhães, Juliana Branco Novo, Patricia Bianca Clissa, Maisa Splendore Della Casa, Diego Butera, Ana Maria Moura da Silva
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  ABSTRACT: Due to its specialized post-translational machinery, mammalian cells represent an interesting and not fully explored system to express snake toxins. Therefore, in this work, we built up a new mammalian expression vector that enhances the feasibility to use mammalian cells to express proteins as biomarkers. Among the modifications, an Igκ signal peptide and a 6xHis tag were inserted into this vector in order to drive the protein to the supernatant and simplify its purification, respectively. In addition, to facilitate selection of high producing clones and also tag proteins which may function as a biomarker, the sequence of enhanced green fluorescent protein (EGFP) was added. The efficiency of the resulting vector (pToxEGFP) was tested by cloning and expressing the viper venom disintegrin echistatin (Ech) that due to its affinity to integrin αvβ3 was tested as a molecular marker. Expression of EGFP-Ech was achieved in CHO-DXB11 cells resulting in a yield of 22 mg/L. The binding activity of this chimera protein was successfully achieved on human umbilical vein endothelial cells which highly express αvβ3. The results indicate that pToxEGFP may constitute an efficient and versatile expression vector to express tagged proteins with potential biomarker activity.
  Molecular Biotechnology 08/2011; 51(2):119-27. · 2.17 Impact Factor
• Source

  Available from: Daniel Carvalho Pimenta

  Article: "Insularin, a disintegrin from Bothrops insularis venom: inhibition of platelet aggregation and endothelial cell adhesion by the native and recombinant GST-insularin proteins".

  Maisa Splendore Della-Casa, Inácio Junqueira-de-Azevedo, Diego Butera, Patrícia Bianca Clissa, Daiana S Lopes, Solange M T Serrano, Daniel C Pimenta, Geraldo S Magalhães, Paulo Lee Ho, Ana Maria Moura-da-Silva
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  ABSTRACT: Insularin (INS) was obtained from Bothrops insularis venom by reversed-phase high-performance liquid chromatography using a C(18) column and characterized as a disintegrin by peptide mass fingerprint and inhibition of ADP-induced platelet aggregation. A cDNA coding for P-II a metalloproteinase/disintegrin was cloned from a cDNA library from B. insularis venom glands. The deduced protein sequence possesses 73 amino acid residues, including the N-terminal, internal peptides of native insularin, the ARGDNP-sequence and 12 cysteines in a conserved alignment. This cDNA fragment was subcloned in the pGEX-4T-1 vector and expressed in a prokaryotic expression system as a fusion protein with glutathione S-transferase (GST-INS). Both native and recombinant insularin inhibited ADP-induced platelet aggregation and endothelial cells (HUVEC) adhesion with similar activities indicating that GST-INS folded correctly and preserved the integrin-binding loop. Insularin may be a tool in studies that involve platelets and endothelial cell adhesion dependent on alphaIIbeta3 and alphavbeta3 integrins.
  Toxicon 11/2010; 57(1):125-33. · 2.51 Impact Factor