Stability improvement of the fatty acid binding protein Sm14 from S. mansoni by Cys replacement: Structural and functional characterization of a vaccine candidate

Celso R R Ramos, Alberto Spisni, Sérgio Oyama, Mauricio L. Sforça, Henrique R. Ramos, Mônica M. Vilar, Adriana C. Alves, Rita C R Figueredo, Míriam Tendler, Nilson I T Zanchin, Thelma A. Pertinhez, Paulo Lee Ho

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The Schistosoma mansoni fatty acid binding protein (FABP), Sm14, is a vaccine candidate against, S. mansoni and F. hepatica. Previously, we demonstrated the importance of a correct fold to achieve protection in immunized animals after cercariae challenge [[10]. C.R.R. Ramos, R.C.R. Figueredo, T.A. Pertinhez, M.M. Vilar, A.L.T.O. Nascimento, M. Tendler, I. Raw, A. Spisni, P.L. Ho, Gene structure and M20T polymorphism of the Schistosoma mansoni Sm14 fatty acid-binding protein: structural, functional and immunoprotection analysis. J. Biol. Chem. 278 (2003) 12745-12751.]. Here we show that the reduction of vaccine efficacy over time is due to protein dimerization and subsequent aggregation. We produced the mutants Sm14-M20(C62S) and Sm14-M20(C62V) that, as expected, did not dimerize in SDS-PAGE. Molecular dynamics calculations and unfolding experiments highlighted a higher structural stability of these mutants with respect to the wild-type. In addition, we found that the mutated proteins, after thermal denaturation, refolded to their active native molecular architecture as proved by the recovery of the fatty acid binding ability. Sm14-M20(C62V) turned out to be the more stable form over time, providing the basis to determine the first 3D solution structure of a Sm14 protein in its apo-form. Overall, Sm14-M20(C62V) possesses an improved structural stability over time, an essential feature to preserve its immunization capability and, in experimentally immunized animals, it exhibits a protection effect against S. mansoni cercariae infections comparable to the one obtained with the wild-type protein. These facts indicate this protein as a good lead molecule for large-scale production and for developing an effective Sm14 based anti-helminthes vaccine.

Original languageEnglish
Pages (from-to)655-662
Number of pages8
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1794
Issue number4
DOIs
Publication statusPublished - Apr 2009

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Fatty Acid-Binding Proteins
Protein S
Schistosoma mansoni
Vaccines
Cercaria
Proteins
Protein Multimerization
Ranunculaceae
Animals
Schistosomiasis mansoni
Immunization
Molecular Dynamics Simulation
Denaturation
Dimerization
Polyacrylamide Gel Electrophoresis
Polymorphism
Fatty Acids
Hot Temperature
Molecular dynamics
Agglomeration

Keywords

  • Fatty-acid binding protein
  • Nuclear Magnetic Resonance
  • Schistosoma mansoni
  • Schistosomiasis
  • Sm14
  • Vaccine

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Analytical Chemistry
  • Molecular Biology

Cite this

Stability improvement of the fatty acid binding protein Sm14 from S. mansoni by Cys replacement : Structural and functional characterization of a vaccine candidate. / Ramos, Celso R R; Spisni, Alberto; Oyama, Sérgio; Sforça, Mauricio L.; Ramos, Henrique R.; Vilar, Mônica M.; Alves, Adriana C.; Figueredo, Rita C R; Tendler, Míriam; Zanchin, Nilson I T; Pertinhez, Thelma A.; Ho, Paulo Lee.

In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1794, No. 4, 04.2009, p. 655-662.

Research output: Contribution to journalArticle

Ramos, CRR, Spisni, A, Oyama, S, Sforça, ML, Ramos, HR, Vilar, MM, Alves, AC, Figueredo, RCR, Tendler, M, Zanchin, NIT, Pertinhez, TA & Ho, PL 2009, 'Stability improvement of the fatty acid binding protein Sm14 from S. mansoni by Cys replacement: Structural and functional characterization of a vaccine candidate', Biochimica et Biophysica Acta - Proteins and Proteomics, vol. 1794, no. 4, pp. 655-662. https://doi.org/10.1016/j.bbapap.2008.12.010
Ramos, Celso R R ; Spisni, Alberto ; Oyama, Sérgio ; Sforça, Mauricio L. ; Ramos, Henrique R. ; Vilar, Mônica M. ; Alves, Adriana C. ; Figueredo, Rita C R ; Tendler, Míriam ; Zanchin, Nilson I T ; Pertinhez, Thelma A. ; Ho, Paulo Lee. / Stability improvement of the fatty acid binding protein Sm14 from S. mansoni by Cys replacement : Structural and functional characterization of a vaccine candidate. In: Biochimica et Biophysica Acta - Proteins and Proteomics. 2009 ; Vol. 1794, No. 4. pp. 655-662.
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