Cratylia mollis Seed Lectin Induces Membrane Permeability Transition in Isolated Rat Liver Mitochondria and a Cyclosporine A‐Insensitive Permeability Transition in Trypanosoma cruzi Mitochondria

• Published in: The Journal of eukaryotic microbiology 2014-07, Vol.61 (4), p.381-388
• Main Authors: Fernandes, Mariana P, Leite, Ana C. R, Araújo, Flavia F. B, Saad, Sara T. O, Baratti, M. O, Correia, M. T. S, Coelho, Luana C. B. B, Gadelha, Fernanda R, Vercesi, Anibal E
• Format: Article
• Language: English
• Subjects: Animals; Biological Transport - drug effects; Calcium; Calcium - metabolism; catalase; confocal microscopy; Cramoll 1; Cramoll 1,4; Cratylia mollis; cyclosporine; Cyclosporine - pharmacology; death; epimastigotes; ethylene glycol tetraacetic acid; Fabaceae - chemistry; glucose; glycoconjugates; hydrogen peroxide; lectins; Lectins - pharmacology; liver; membrane permeability; membrane potential; Membrane Potential, Mitochondrial - drug effects; mitochondria; Mitochondria - drug effects; Mitochondria - metabolism; Mitochondria, Liver - drug effects; Mitochondria, Liver - metabolism; mitochondrial membrane; mitochondrial permeability transition; necrosis; plasma membrane; Rats; Reactive Oxygen Species - metabolism; ROS; Seeds - chemistry; Trypanosoma cruzi; Trypanosoma cruzi - drug effects; Trypanosoma cruzi - metabolism
• ISSN: 1066-5234; 1550-7408
• DOI: 10.1111/jeu.12118

Previous results provided evidence that Cratylia mollis seed lectin (Cramoll 1,4) promotes Trypanosoma cruzi epimastigotes death by necrosis via a mechanism involving plasma membrane permeabilization to Ca²⁺and mitochondrial dysfunction due to matrix Ca²⁺overload. In order to investigate the mechanism of Ca²⁺‐induced mitochondrial impairment, experiments were performed analyzing the effects of this lectin on T. cruzi mitochondrial fraction and in isolated rat liver mitochondria (RLM), as a control. Confocal microscopy of T. cruzi whole cell revealed that Cramoll 1,4 binding to the plasma membrane glycoconjugates is followed by its internalization and binding to the mitochondrion. Electrical membrane potential (∆Ψₘ) of T. cruzi mitochondrial fraction suspended in a reaction medium containing 10 μM Ca²⁺was significantly decreased by 50 μg/ml Cramoll 1,4 via a mechanism insensitive to cyclosporine A (CsA, membrane permeability transition (MPT) inhibitor), but sensitive to catalase or 125 mM glucose. In RLM suspended in a medium containing 10 μM Ca²⁺this lectin, at 50 μg/ml, induced increase in the rate of hydrogen peroxide release, mitochondrial swelling, and ∆Ψₘdisruption. All these mitochondrial alterations were sensitive to CsA, catalase, and EGTA. These results indicate that Cramoll 1, 4 leads to inner mitochondrial membrane permeabilization through Ca²⁺dependent mechanisms in both mitochondria. The sensitivity to CsA in RLM characterizes this lectin as a MPT inducer and the lack of CsA effect identifies a CsA‐insensitive MPT in T. cruzi mitochondria.