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Memórias do Instituto Oswaldo Cruz
Fundação Oswaldo Cruz, Fiocruz
ISSN: 1678-8060
EISSN: 1678-8060
Vol. 107, No. 6, 2012, pp. 752-759
Bioline Code: oc12117
Full paper language: English
Document type: Research Article
Document available free of charge

Memórias do Instituto Oswaldo Cruz, Vol. 107, No. 6, 2012, pp. 752-759

 en Proteolytic activity in the adult and larval stages of the human roundworm parasite Angiostrongylus costaricensis check for this species in other resources
Rebello, Karina Mastropasqua; de Siqueira, Caroline Reis; Ribeiro, Erika Louise; Valente, Richard Hemmi; Mota, Ester Maria; Perales, Jonas; Neves-Ferreira, Ana Gisele da Costa & Lenzi, Henrique Leonel

Abstract

Angiostrongylus costaricensis check for this species in other resources is a nematode that causes abdominal angiostrongyliasis, a widespread human parasitism in Latin America. This study aimed to characterize the protease profiles of different developmental stages of this helminth. First-stage larvae (L1) were obtained from the faeces of infected Sigmodon hispidus check for this species in other resources rodents and third-stage larvae (L3) were collected from mollusks Biomphalaria glabrata check for this species in other resources previously infected with L1. Adult worms were recovered from rodent mesenteric arteries. Protein extraction was performed after repeated freeze-thaw cycles followed by maceration of the nematodes in 40 mM Tris base. Proteolysis of gelatin was observed by zymography and found only in the larval stages. In L3, the gelatinolytic activity was effectively inhibited by orthophenanthroline, indicating the involvement of metalloproteases. The mechanistic class of the gelatinases from L1 could not be precisely determined using traditional class-specific inhibitors. Adult worm extracts were able to hydrolyze haemoglobin in solution, although no activity was observed by zymography. This haemoglobinolytic activity was ascribed to aspartic proteases following its effective inhibition by pepstatin, which also inhibited the haemoglobinolytic activity of L1 and L3 extracts. The characterization of protease expression throughout the A. costaricensis life cycle may reveal key factors influencing the process of parasitic infection and thus foster our understanding of the disease pathogenesis.

Keywords
Angiostrongylus costaricensis - metalloproteases - aspartic proteases - haemoglobin - zymography

 
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