Apparent isocitrate lyase activity in Leishmania amazonensis

Concepción Hernández-Chinea; Laura Maimone; Yelitza Campos; Walter Mosca; Pedro J. Romero

doi:10.1515/ap-2017-0084

Artikel

Apparent isocitrate lyase activity in Leishmania amazonensis

Concepción Hernández-Chinea , Laura Maimone , Yelitza Campos , Walter Mosca und Pedro J. Romero

Veröffentlicht/Copyright: 15. Oktober 2017

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Aus der Zeitschrift Acta Parasitologica Band 62 Heft 4

Abstract

Early reports have demonstrated the occurrence of glyoxylate cycle enzymes in several Leishmania species. However, these results have been underestimated because genes for the two key enzymes of the cycle, isocitrate lyase (ICL) and malate synthase (MS), are not annotated in Leishmania genomes. We have re-examined this issue in promastigotes of Leishmania amazonensis. Enzyme activities were assayed spectrophotometrically in cellular extracts and characterized partially. A 40 kDa band displaying ICL activity was visualized on zymograms of the extracts. By immunoblotting with mouse antibodies against ICL from Bacillus stearothermophilus, a band of approximately 40 kDa was identified, coincident with the relative molecular mass of the activity band revealed on zymograms. Indirect immunofluorescence of intact promastigotes showed that the recognized antigen is distributed as a punctuated pattern, mainly distributed beneath the subpellicular microtubules, over a diffused cytoplasmic stain. These results clearly demonstrate the existence of an apparent ICL activity in L. amazonensis promastigotes, which is associated to a 40 kDa polypeptide and distributed both diffused and as punctuate aggregates in the cytoplasm. The relevance of this activity is discussed.

Keywords: Leishmania amazonensis; promastigote; glyoxylate cycle; isocitrate lyase; malate synthase

Acknowledgements

This work was financially supported by Fondo Nacional de Ciencia y Tecnología (FONACIT), [grant number S1-2001000668] and Consejo de Desarrollo Científico y Humanístico de la Universidad Central de Venezuela (CDCH-UCV), [grant number PI 03-33-5462-2004] to C.H-Ch.

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Received: 2016-8-15

Revised: 2017-2-22

Accepted: 2017-6-23

Published Online: 2017-10-15

Published in Print: 2017-12-20

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https://doi.org/10.1515/ap-2017-0084

Schlagwörter für diesen Artikel

Leishmania amazonensis; promastigote; glyoxylate cycle; isocitrate lyase; malate synthase