, Volume 179, Issue 3–4, pp 231–242 | Cite as

1,10-Phenanthroline Inhibits the Metallopeptidase Secreted by Phialophora verrucosa and Modulates its Growth, Morphology and Differentiation

  • Marcela Queiroz Granato
  • Priscila de Araújo Massapust
  • Sonia Rozental
  • Celuta Sales Alviano
  • André Luis Souza dos Santos
  • Lucimar Ferreira Kneipp


Phialophora verrucosa is one of the etiologic agents of chromoblastomycosis, a fungal infection that affects cutaneous and subcutaneous tissues. This disease is chronic, recurrent and difficult to treat. Several studies have shown that secreted peptidases by fungi are associated with important pathophysiological processes. Herein, we have identified and partially characterized the peptidase activity secreted by P. verrucosa conidial cells. Using human serum albumin as substrate, the best hydrolysis profile was detected at extreme acidic pH (3.0) and at 37 °C. The enzymatic activity was completely blocked by classical metallopeptidase inhibitors/chelating agents as 1,10-phenanthroline and EGTA. Zinc ions stimulated the metallo-type peptidase activity in a dose-dependent manner. Several proteinaceous substrates were cleaved, in different extension, by the P. verrucosa metallopeptidase activity, including immunoglobulin G, fibrinogen, collagen types I and IV, fibronectin, laminin and keratin; however, mucin and hemoglobin were not susceptible to proteolysis. As metallopeptidases participate in different cellular metabolic pathways in fungal cells, we also tested the influence of 1,10-phenanthroline and EGTA on P. verrucosa development. Contrarily to EGTA, 1,10-phenanthroline inhibited the fungal viability (MIC 0.8 µg/ml), showing fungistatic effect, and induced profound morphological alterations as visualized by transmission electron microscopy. In addition, 1,10-phenanthroline arrested the filamentation process in P. verrucosa. Our results corroborate the supposition that metallopeptidase inhibitors/chelating agents have potential to control crucial biological events in fungal agents of chromoblastomycosis.


Phialophora verrucosa Chromoblastomycosis Metallopeptidase 1,10-Phenanthroline Growth Differentiation 



This work was supported mainly by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). This study was also financed by Grants from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) and Fundação Oswaldo Cruz (FIOCRUZ).

Conflict of interest



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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Marcela Queiroz Granato
    • 1
  • Priscila de Araújo Massapust
    • 1
  • Sonia Rozental
    • 2
  • Celuta Sales Alviano
    • 3
  • André Luis Souza dos Santos
    • 4
    • 5
  • Lucimar Ferreira Kneipp
    • 1
  1. 1.Laboratório de Taxonomia, Bioquímica e Bioprospecção de Fungos, Instituto Oswaldo CruzFundação Oswaldo CruzRio de JaneiroBrazil
  2. 2.Laboratório de Biologia Celular de Fungos, Instituto de Biofísica Carlos Chagas FilhoUniversidade Federal do Rio de Janeiro (UFRJ)Rio de JaneiroBrazil
  3. 3.Laboratório de Superfície de Microrganismos, Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de GóesUniversidade Federal do Rio de Janeiro (UFRJ)Rio de JaneiroBrazil
  4. 4.Laboratório de Investigação de Peptidases, Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de GóesUniversidade Federal do Rio de Janeiro (UFRJ)Rio de JaneiroBrazil
  5. 5.Programa de Pós-Graduação em Bioquímica, Instituto de QuímicaUniversidade Federal do Rio de Janeiro (UFRJ)Rio de JaneiroBrazil

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