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Leishmania mexicana: identification and characterization of an aspartyl proteinase activity.

Experimental parasitology (2006-11-28)
Elizabeth Valdivieso, Fracehuli Dagger, Ana Rascón
ABSTRACT

An aspartyl proteinase activity was detected in the soluble fraction (SF) of Leishmania mexicana promastigotes by the use of the synthetic substrate benzoyl-Arg-Gly-Phe-Phe-Leu-4-methoxy-beta-naphthylamide selective for Cathepsin D like aspartyl-proteinases. This peptide was hydrolyzed with an apparent K(m) of 2.3+/-0.3 microM. The classic inhibitor of aspartyl-proteinases, diazo-acetyl-norleucinemethylester (DAN) inhibited the proteolytic activity with an IC(50) of 400 microM. The soluble fraction degraded (in absence of thiol groups) human fibrinogen with a specific activity of 533 U/mg protein. When tested for the ability to inhibit the "in vitro" proliferation of L. mexicana promastigotes, DAN showed concentration dependent anti-proliferative effects with a LD(50) of 466 microM at 48 h, with a significant fall in this value to 22 microM after 72 h. This is the first characterization of an aspartyl-proteinase activity in Leishmania, calling for further studies directed towards the physiologic role of these enzymes in the parasite. The anti-proliferative effect of its inhibition makes this enzyme a putative new target for the development of leishmanicidal drugs.