Lytic activity and structural differences of amphipathic peptides derived from trialysin

Rafael M. Martins, Maurício L. Sforça, Rogério Amino, Maria Aparecida Juliano, Sérgio Oyama, Luiz Juliano, Thelma A. Pertinhez, Alberto Spisni, Sergio Schenkman

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Trialysin is a pore-forming protein found in the saliva of Triatoma infestans (Hemiptera, Reduviidae), the insect vector of Chagas' disease. The protein is active against a broad range of cell types from bacteria to eukaryotic cells. Recognizing that the N-terminus of trialysin harbors the lytic motif [Amino, R., Martins, R. M., Procopio, J., Hirata, I. Y., Juliano, M. A., and Schenkman, S. (2002) J. Biol. Chem. 277, 6207-6213], we designed a set of peptides scanning this region to investigate the structural basis of its biological function. Peptides encompassing residues 1-32 (P6), 1-27 (P7), and 6-32 (P5) efficiently induced lysis of the protozoan parasite Trypanosoma cruzi and Escherichia coli in the 0.4-9.0 μM range, while much higher concentrations were required to cause hemolysis. Other more internal peptides, including peptide P2 (residues 21-47) and others up to residue 52, were less effective. P6 turned out to be the most active of all. P7 has a significantly higher activity than P5 against E. coli, while P5 has a hemolytic activity comparable to that of P6. CD spectroscopy showed that all tested peptides acquire a comparable helical content in solvent mixtures or in detergent micelles. The solution structure of P2 and P5-P7 was determined in a 30% trifluoroethanol/water mixture by nuclear magnetic resonance. All peptides exhibit a structure characterized by a central helical fold, and except for P2, which does not show a continuous hydrophobic surface, they are amphipathic. The structural models show that P5 and P7 extend their structural similarities with the most active peptide, P6, in either the C-terminus or the N-terminus. Amino acid substitutions in the N-terminus of P6 improved hemolysis but did not change the activity against T. cruzi. These results suggest that while amphipathicity is essential for the lytic activity, the selectivity of the active peptides for specific organisms appears to be associated with the structural features of their N- and C-termini.

Original languageEnglish
Pages (from-to)1765-1774
Number of pages10
JournalBiochemistry
Volume45
Issue number6
DOIs
Publication statusPublished - Feb 14 2006

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Peptides
Trypanosoma cruzi
Hemolysis
Escherichia coli
Reduviidae
Insect Vectors
Triatoma
Trifluoroethanol
Porins
Hemiptera
Chagas Disease
Structural Models
Micelles
Eukaryotic Cells
Amino Acid Substitution
Ports and harbors
Saliva
Detergents
Spectrum Analysis
Bacteria

ASJC Scopus subject areas

  • Biochemistry

Cite this

Martins, R. M., Sforça, M. L., Amino, R., Juliano, M. A., Oyama, S., Juliano, L., ... Schenkman, S. (2006). Lytic activity and structural differences of amphipathic peptides derived from trialysin. Biochemistry, 45(6), 1765-1774. https://doi.org/10.1021/bi0514515

Lytic activity and structural differences of amphipathic peptides derived from trialysin. / Martins, Rafael M.; Sforça, Maurício L.; Amino, Rogério; Juliano, Maria Aparecida; Oyama, Sérgio; Juliano, Luiz; Pertinhez, Thelma A.; Spisni, Alberto; Schenkman, Sergio.

In: Biochemistry, Vol. 45, No. 6, 14.02.2006, p. 1765-1774.

Research output: Contribution to journalArticle

Martins, RM, Sforça, ML, Amino, R, Juliano, MA, Oyama, S, Juliano, L, Pertinhez, TA, Spisni, A & Schenkman, S 2006, 'Lytic activity and structural differences of amphipathic peptides derived from trialysin', Biochemistry, vol. 45, no. 6, pp. 1765-1774. https://doi.org/10.1021/bi0514515
Martins RM, Sforça ML, Amino R, Juliano MA, Oyama S, Juliano L et al. Lytic activity and structural differences of amphipathic peptides derived from trialysin. Biochemistry. 2006 Feb 14;45(6):1765-1774. https://doi.org/10.1021/bi0514515
Martins, Rafael M. ; Sforça, Maurício L. ; Amino, Rogério ; Juliano, Maria Aparecida ; Oyama, Sérgio ; Juliano, Luiz ; Pertinhez, Thelma A. ; Spisni, Alberto ; Schenkman, Sergio. / Lytic activity and structural differences of amphipathic peptides derived from trialysin. In: Biochemistry. 2006 ; Vol. 45, No. 6. pp. 1765-1774.
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abstract = "Trialysin is a pore-forming protein found in the saliva of Triatoma infestans (Hemiptera, Reduviidae), the insect vector of Chagas' disease. The protein is active against a broad range of cell types from bacteria to eukaryotic cells. Recognizing that the N-terminus of trialysin harbors the lytic motif [Amino, R., Martins, R. M., Procopio, J., Hirata, I. Y., Juliano, M. A., and Schenkman, S. (2002) J. Biol. Chem. 277, 6207-6213], we designed a set of peptides scanning this region to investigate the structural basis of its biological function. Peptides encompassing residues 1-32 (P6), 1-27 (P7), and 6-32 (P5) efficiently induced lysis of the protozoan parasite Trypanosoma cruzi and Escherichia coli in the 0.4-9.0 μM range, while much higher concentrations were required to cause hemolysis. Other more internal peptides, including peptide P2 (residues 21-47) and others up to residue 52, were less effective. P6 turned out to be the most active of all. P7 has a significantly higher activity than P5 against E. coli, while P5 has a hemolytic activity comparable to that of P6. CD spectroscopy showed that all tested peptides acquire a comparable helical content in solvent mixtures or in detergent micelles. The solution structure of P2 and P5-P7 was determined in a 30{\%} trifluoroethanol/water mixture by nuclear magnetic resonance. All peptides exhibit a structure characterized by a central helical fold, and except for P2, which does not show a continuous hydrophobic surface, they are amphipathic. The structural models show that P5 and P7 extend their structural similarities with the most active peptide, P6, in either the C-terminus or the N-terminus. Amino acid substitutions in the N-terminus of P6 improved hemolysis but did not change the activity against T. cruzi. These results suggest that while amphipathicity is essential for the lytic activity, the selectivity of the active peptides for specific organisms appears to be associated with the structural features of their N- and C-termini.",
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AU - Oyama, Sérgio

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N2 - Trialysin is a pore-forming protein found in the saliva of Triatoma infestans (Hemiptera, Reduviidae), the insect vector of Chagas' disease. The protein is active against a broad range of cell types from bacteria to eukaryotic cells. Recognizing that the N-terminus of trialysin harbors the lytic motif [Amino, R., Martins, R. M., Procopio, J., Hirata, I. Y., Juliano, M. A., and Schenkman, S. (2002) J. Biol. Chem. 277, 6207-6213], we designed a set of peptides scanning this region to investigate the structural basis of its biological function. Peptides encompassing residues 1-32 (P6), 1-27 (P7), and 6-32 (P5) efficiently induced lysis of the protozoan parasite Trypanosoma cruzi and Escherichia coli in the 0.4-9.0 μM range, while much higher concentrations were required to cause hemolysis. Other more internal peptides, including peptide P2 (residues 21-47) and others up to residue 52, were less effective. P6 turned out to be the most active of all. P7 has a significantly higher activity than P5 against E. coli, while P5 has a hemolytic activity comparable to that of P6. CD spectroscopy showed that all tested peptides acquire a comparable helical content in solvent mixtures or in detergent micelles. The solution structure of P2 and P5-P7 was determined in a 30% trifluoroethanol/water mixture by nuclear magnetic resonance. All peptides exhibit a structure characterized by a central helical fold, and except for P2, which does not show a continuous hydrophobic surface, they are amphipathic. The structural models show that P5 and P7 extend their structural similarities with the most active peptide, P6, in either the C-terminus or the N-terminus. Amino acid substitutions in the N-terminus of P6 improved hemolysis but did not change the activity against T. cruzi. These results suggest that while amphipathicity is essential for the lytic activity, the selectivity of the active peptides for specific organisms appears to be associated with the structural features of their N- and C-termini.

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