The protozoan parasite Entamoeba histolytica is an ancient eukaryotic cell that shows morphologically atypical organelles and differs metabolically from higher eukaryotic cells. The aim of this study was to determine the subcellular localization of ameba NAD+-dependent alcohol dehydrogenase (ADH2). The enzyme activity was present in soluble and mainly in particulate material whose density was 1.105 in a sucrose gradient. By differential centrifugation, most of the ADH activity sedimented at 160,000 g (160,000-g pellet), similar to the Escherichia coli polymeric ADHE. In the Coomassie staining of the 160,000-g pellet analyzed by electrophoresis, a 96-kDa protein was more prominent than in other fractions; this band was recognized by antibodies against Lactococcus lactis ADHE. By gold labeling, the antibodies recognized the granular material that mainly constitutes the 160,000-g pellet and a material that sedimented along with the internal membrane vesicles. By negative staining, the 160,000-g fraction showed helical rodlike structures with an average length of 103 nm; almost no membrane vesicles were observed in this pellet. In internal membrane fractions, no rodlike structures were found, but protomerlike round structures were observed. These results indicate that the main amebic NAD+-dependent ADH2 activity is naturally organized as rodlike helical particles, similar to bacterial ADHE. Detection of ADH2 in membrane fractions might be explained by cosedimentation of the multimeric ADH during membrane purification.
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April 2002
BIOCHEMISTRY-PHYSIOLOGY|
April 01 2002
SUBCELLULAR LOCALIZATION OF THE NAD+-DEPENDENT ALCOHOL DEHYDROGENASE IN ENTAMOEBA HISTOLYTICA TROPHOZOITES
Eva E. Avila;
Eva E. Avila
Instituto de Investigación en Biología Experimental, Facultad de Química, Universidad de Guanajuato. P.O. Box 187, Guanajuato, Gto. México. [email protected]
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Edith R. Martínez-Alcaraz;
Edith R. Martínez-Alcaraz
Instituto de Investigación en Biología Experimental, Facultad de Química, Universidad de Guanajuato. P.O. Box 187, Guanajuato, Gto. México. [email protected]
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Gloria Barbosa-Sabanero;
Gloria Barbosa-Sabanero
Instituto de Investigación en Biología Experimental, Facultad de Química, Universidad de Guanajuato. P.O. Box 187, Guanajuato, Gto. México. [email protected]
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Elda I. Rivera-Baron;
Elda I. Rivera-Baron
Instituto de Investigación en Biología Experimental, Facultad de Química, Universidad de Guanajuato. P.O. Box 187, Guanajuato, Gto. México. [email protected]
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Sergio Arias-Negrete;
Sergio Arias-Negrete
Instituto de Investigación en Biología Experimental, Facultad de Química, Universidad de Guanajuato. P.O. Box 187, Guanajuato, Gto. México. [email protected]
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Roberto Zazueta-Sandoval
Roberto Zazueta-Sandoval
Instituto de Investigación en Biología Experimental, Facultad de Química, Universidad de Guanajuato. P.O. Box 187, Guanajuato, Gto. México. [email protected]
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J Parasitol (2002) 88 (2): 217–222.
Citation
Eva E. Avila, Edith R. Martínez-Alcaraz, Gloria Barbosa-Sabanero, Elda I. Rivera-Baron, Sergio Arias-Negrete, Roberto Zazueta-Sandoval; SUBCELLULAR LOCALIZATION OF THE NAD+-DEPENDENT ALCOHOL DEHYDROGENASE IN ENTAMOEBA HISTOLYTICA TROPHOZOITES. J Parasitol 1 April 2002; 88 (2): 217–222. doi: https://doi.org/10.1645/0022-3395(2002)088[0217:SLOTND]2.0.CO;2
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