Proteinase activities during temperature-induced stage differentiation of species complexes of Leishmania.
We have examined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), using gelatin, bovine serum albumin (BSA) or human IgG as substrate, proteinase activities in cell lysates from selected species complexes of Leishmania. The inhibition of proteinase activity caused by the reagent L-trans-epoxysuccinylleucylamido(4-guanidino)butane (E-64), which is known to act only on cysteinyl proteinases, revealed a 31 kDa component of this class of enzymes in soluble, but not in membrane-enriched preparations, of either L. amazonensis or L. major-like parasites from the New World. The proteinase component was detectable in the leishmanial multiplicative promastigote stage (log phase) and its concentration apparently increased during the thermally induced transformation of promastigotes to amastigote-like forms in vitro. Comparative studies revealed that taxonomically distinct species complexes of Leishmania possess high amastigote cysteine proteinase activity. This feature, however, was lacking in other developmental stages of the species (L. braziliensis, L. chagasi, L. aethiopica, and L. donovani) analyzed. Furthermore, lesion amastigotes of L. amazonensis displayed ultrastructurally recognizable megasomes, but megasome-like or large multivesicular body organelles could be detected only in axenic amastigotes of both L. amazonensis and L. major-like species.
Leon
L L
LL
Departamento de Imunologia, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil.
Temporal
R M
RM
Soares
M J
MJ
Grimaldi Júnior
G
G
D000818:Animals; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D006801:Humans; D007891:Leishmania / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D013045:Species Specificity; D013379:Substrate Specificity
D015853:Cysteine Proteinase Inhibitors; D003546:Cysteine Endopeptidases; D007930:Leucine; C024974:E 64