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Detection of cysteine-proteinases in Leishmania amazonensis promastigotes using a cross-reactive antiserum
Authors:Alves C R  Côrte-Real S  De-Freitas Rosa M  Giovanni-De-Simone S
Affiliation:Departamento de Bioquímica e Biologia Molecular, Instituto Oswaldo Cruz, FIOCRUZ, CP 926, 21045-900, Rio de Janeiro, RJ, Brazil. calves@gene.dbbm.fiocruz.br
Abstract:Rabbit serum against the cysteine-proteinases papain has been employed for the cellular localization of cysteine-proteinases of in Leishmania amazonensis promastigotes. By immunocytochemistry, immune complexes were found in the plasma membrane and in the flagella pocket of the parasite. The antiserum immunoprecipitated major iodinated proteins with molecular masses of 66, 45, 28 and 24 kDa and a wide partitioning of the Triton X-114 detergent phase. The presence of cysteine-proteinase at the cell surface membrane was also suggested by the detection of proteolytic activity in living cells (19.0 microg azocasein min(-1) 10(-7) promastigotes (1.0 S.D. )).
Keywords:Leishmania amazonensis    Cysteine-proteinase    Anti-papain serum    Immunocytochemistry
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