A putative calcium-ATPase of the secretory pathway family may regulate calcium/manganese levels in the Golgi apparatus of Entamoeba histolytica
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Calcium regulates many cellular processes in protozoa, including growth, differentiation, programmed cell death, exocytosis, endocytosis, phagocytosis, fusion of the endosomes of distinct stages with phagosomes, fusion of phagosomes with lysosomes, and recycling the membrane. In Entamoeba histolytica, the protozoa responsible for human amoebiasis, calcium ions are essential for signaling pathways that lead to growth and development. In addition, calcium is crucial in the modulation of gene expression in this microorganism. However, there is scant information about the proteins responsible for regulating calcium levels in this parasite. In this work, we characterized a protein of E. histolytica that shows a close phylogenetic relationship with Ca2+ pumps that belong to the family of secretory pathway calcium ATPases (SPCA), which for several organisms are located in the Golgi apparatus. The amoeba protein analyzed herein has several amino acid residues that are characteristic of SPCA members. By an immunofluorescent technique using specific antibodies and immunoelectron microscopy, the protein was detected on the membrane of some cytoplasmic vacuoles. Moreover, this putative calcium-ATPase was located in vacuoles stained with NBD C6-ceramide, a Golgi marker. Overall, the current findings support the hypothesis that the presently analyzed protein corresponds to the SPCA of E. histolytica.
KeywordsEntamoeba histolytica Golgi apparatus Calcium-ATPases SPCA
We thank to Carlos Vázquez-Calzada, for his invaluable technical assistance (Centro de Investigación y de Estudios Avanzados del IPN).
Compliance with ethical standards
Conflict of Interest
The authors declare that they have no conflict of interest.
- Cohen Y, Megyeri M, Chen OCW, Condomitti G, Riezman I, Loizides-Mangold U, Abdul-Sada A, Rimon N, Riezman H, Platt FM, Futerman AH, Schuldiner M (2013) The yeast p5 type ATPase, spf1, regulates manganese transport into the endoplasmic reticulum. PLoS One 8(12):e85519. https://doi.org/10.1371/journal.pone.0085519 CrossRefPubMedPubMedCentralGoogle Scholar
- Dode L, Andersen JP, Vanoevelen J, Raeymaekers L, Missiaen L, Vilsen B, Wuytack F (2006) Dissection of the functional differences between human secretory pathway Ca2+/Mn2+-ATPase (SPCA) 1 and 2 isoenzymes by steady-state and transient kinetic analyses. J Biol Chem 281(6):3182–3189. https://doi.org/10.1074/jbc.M511547200 CrossRefPubMedGoogle Scholar
- Ghosh SK, Field J, Frisardi M, Rosenthal B, Mai Z, Rogers R, Samuelson J (1999) Chitinase secretion by encysting Entamoeba invadens and transfected Entamoeba histolytica trophozoites: localization of secretory vesicles, endoplasmic reticulum, and Golgi apparatus. Infect Immun 67(6):3073–3081PubMedPubMedCentralGoogle Scholar
- Jain R, Santi-Rocca J, Padhan N, Bhattacharya S, Guillen N, Bhattacharya A (2008) Calcium-binding protein 1 of Entamoeba histolytica transiently associates with phagocytic cups in a calcium-independent manner. Cell Microbiol 10(6):1373–1389. https://doi.org/10.1111/j.1462-5822.2008.01134.x CrossRefPubMedGoogle Scholar
- Kumar S, Aslam S, Mazumder M, Dahiya P, Murmu A, Manjasetty BA, Zaidi R, Bhattacharya A, Gourinath S (2014) Crystal structure of calcium binding protein-5 from Entamoeba histolytica and its involvement in initiation of phagocytosis of human erythrocytes. PLoS Pathog 10(12):e1004532. https://doi.org/10.1371/journal.ppat.1004532 CrossRefPubMedPubMedCentralGoogle Scholar
- Martinez-Higuera A, Salas-Casas A, Calixto-Gálvez M, Chávez-Munguía B, Pérez-Ishiwara DG, Ximénez C, Rodríguez MA (2013) Identification of calcium-transporting ATPases of Entamoeba histolytica and cellular localization of the putative SERCA. Exp Parasitol 135(1):79–86. https://doi.org/10.1016/j.exppara.2013.06.004 CrossRefPubMedGoogle Scholar
- Mitchell KJ, Pinton P, Varadi A, Tacchetti C, Ainscow EK, Pozzan T, Rizzuto R, Rutter GA (2001) Dense core secretory vesicles revealed as a dynamic Ca(2+) store in neuroendocrine cells with a vesicle-associated membrane protein aequorin chimaera. J Cell Biol 155(1):41–51. https://doi.org/10.1083/jcb.200103145 CrossRefPubMedPubMedCentralGoogle Scholar
- Segovia-Gamboa NC, Talamás-Rohana P, Ángel-Martínez A, Cázares-Raga FE, González-Robles A, Hernández-Ramírez VI, Martínez-Palomo A, Chávez-Munguía B (2011) Differentiation of Entamoeba histolytica: a possible role for enolase. Exp Parasitol 129(1):65–71. https://doi.org/10.1016/j.exppara.2011.05.001 CrossRefPubMedGoogle Scholar
- Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28(10):2731–2739. https://doi.org/10.1093/molbev/msr121 CrossRefPubMedPubMedCentralGoogle Scholar
- WHO (1997) Amoebiasis WHO epidemiol weekly record. vol 72, p 97–100Google Scholar