The pyruvate:ferredoxin oxidoreductase enzyme is located in the plasma membrane and in a cytoplasmic structure inEntamoeba |
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| Institution: | 1. Departamento de Patologı́a Experimental, Centro de Investigación y de Estudios avanzados del IPN, Apdo Postal 14-740, México, 07300, D.F. México;2. Programa Institucional de Biomedicina Molecular, CICATA-IPN, Legaria 694 col. Irrigación, México, 11500, D.F. México;3. Unité de Pathogénie Moléculaire, Institut Pasteur, 28 rue du Dr. Roux F. Paris Cedex 15, 75724, France;1. Institute for Research in Molecular Medicine (INFORMM), Universti Sains Malaysia, Penang, 11800, Malaysia;2. EMAN Testing and Research Laboratories, Department of Pharmacology, School of Pharmaceutical Sciences, Universti Sains Malaysia, Penang 11800, Malaysia;3. ACRF Department of Cancer Biology and Therapeutics, The John Curtin School of Medical Research, Australian National University, Australia;1. University of Miami, School of Medicine, Department of Public Health Sciences, United States;2. Rand Corporation, Santa Monica, CA, United States;3. University of California, San Francisco School of Nursing, Department of Community Health Systems, United States;4. University of California, San Francisco, Center for AIDS Prevention Studies, United States;5. Georgia State University, Department of Psychology, United States;6. Centers for Disease Control and Prevention, United States;1. School of Biological Sciences, University of Queensland, St. Lucia, QLD, 4072, Australia;2. Faculty of Biology, Hanoi National University of Education, Hanoi, Viet Nam;3. Department of Agriculture and Fisheries, Ecosciences Precinct, GPO Box 267, Brisbane, QLD, 4001, Australia;1. School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, 325035, PR China;2. Department of Pharmacology and Toxicology, University of Louisville, Louisville, KY, USA;3. Department of Zoology and Fisheries, University of Agriculture Faisalabad, 38000, Pakistan;4. Agricultural Products Quality and Safety Supervision and Management Bureau, Xuancheng, 242000, PR China;5. Research Institute of Applied Biology, Shanxi University, Taiyuan, 030006, PR China |
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| Abstract: | This work investigated the cellular location of the pyruvate:ferredoxin oxidoreductase (PFO) enzyme inEntamoeba. A 1.9 kb fragment located at the 3 end of theEhpfogene was cloned in the pRSETB vector and expressed. The recombinant peptide was purified and inoculated in rabbits. By Western blot assays the antibodies detected a single 130 kDa band in allE. histolyticastrains tested and inE. moshkovskii. By immunofluorescence, the antibodies showed the presence of PFO in the plasma membrane and in a cytoplasmic structure that appeared as a ring or as a compact small body inE. histolyticastrains. InE. invadensandE. moshkovskii(strains FIC and Laredo) PFO was located in the plasma membrane showing different fluorescence patterns. Immunofluorescence onE. histolyticasynchronized cultures showed that the cytoplasmic structure appeared in 85, 60, 20 and 10% of the trophozoites in mitosis, G1, S and G2 phases, respectively. Byin situhibridization theEhpfogene was found in the nuclei and the trophozoites of the clone A, strain HM1:IMSS, differed in theEhpfogene content. |
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