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Differential effects of α-helical and β-hairpin antimicrobial peptides against Acanthamoeba castellanii

Published online by Cambridge University Press:  02 June 2009

R. S. SACRAMENTO ,
R. M. MARTINS ,
A. MIRANDA ,
A. S. S. DOBROFF ,
S. DAFFRE ,
A. S. FORONDA ,
D. DE FREITAS  and
S. SCHENKMAN
R. S. SACRAMENTO
Affiliation:
Departamento de Oftalmologia, Universidade Federal de São Paulo, SP, Brazil
R. M. MARTINS
Affiliation:
Departamento de Microbiologia, Imunologia, Parasitologia, Universidade Federal de São Paulo, SP, Brazil
A. MIRANDA
Affiliation:
Departamento de Biofísica, Universidade Federal de São Paulo, SP, Brazil
A. S. S. DOBROFF
Affiliation:
Departamento de Microbiologia, Imunologia, Parasitologia, Universidade Federal de São Paulo, SP, Brazil
S. DAFFRE
Affiliation:
Departamento de Parasitologia, Universidade São Paulo, São Paulo, SP, Brazil
A. S. FORONDA
Affiliation:
Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade São Paulo, São Paulo, SP, Brazil
D. DE FREITAS
Affiliation:
Departamento de Oftalmologia, Universidade Federal de São Paulo, SP, Brazil
S. SCHENKMAN*
Affiliation:
Departamento de Microbiologia, Imunologia, Parasitologia, Universidade Federal de São Paulo, SP, Brazil
*
*Corresponding author: Departamento de Microbiologia, Imunologia e Parasitologia, UNIFESP, Rua Botucatu 862, 8A–04023-062, São Paulo, SP, Brazil. Tel: +5511 55751996. E-mail: sschenkman@unifesp.br
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Summary

In this work we evaluated the ability of different types of antimicrobial peptides to promote permeabilization and growth inhibition of Acanthamoeba castellanii trophozoites, which cause eye keratitis. We used cationic α-helical peptides P5 and P6, corresponding to the N-terminus of the pore-forming protein from Triatoma infestans, a blood-sucking insect, and a β-hairpin amphipathic molecule (gomesin), of the spider Acanthoscurria gomesiana haemocytes. A. castellanii permeabilization was obtained after 1 h incubation with micromolar concentrations of both types of peptides. While permeabilization induced by gomesin increased with longer incubations, P5 permeabilization did not increase with time and occurred at doses that are more toxic for SIRC cells. P5, however, at doses below the critical dose used to kill rabbit corneal cells was quite effective in promoting growth inhibition. Similarly, P5 was more effective when serine protease inhibitor was added simultaneously to the permeabilization assay. High performance chromatography followed by mass spectrometry analysis confirmed that, in contrast to gomesin, P5 is hydrolysed by A. castellanii culture supernatants. We conclude that the use of antimicrobial peptides to treat A. castellanii infections requires the search of more specific peptides that are resistant to proteolysis.

Keywords

Acanthamoeba castellaniiantimicrobial peptidesamphipathic peptides permeabilizationprotease
Type
Research Article
Information
Parasitology , Volume 136 , Issue 8 , July 2009 , pp. 813 - 821
Copyright
Copyright © Cambridge University Press 2009

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