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Structural and Biochemical Analyses of Alcohol Dehydrogenase E Enzymes from Entamoeba invadens IP‐1, E. invadens VK‐1:NS and E. dispar
Author(s) -
Gabrielle Matthew,
Leito Joshua,
Espinosa Avelina
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.606.16
Subject(s) - entamoeba histolytica , alcohol dehydrogenase , entamoeba , aldehyde dehydrogenase , biology , enzyme , biochemistry , acetaldehyde , gene , microbiology and biotechnology , ethanol
The Entamoeba lineage belongs to the Amoebozoa, one of six major divisions of eukaryotes. Entamoeba trophozoites, similar to other protists, actively capture ingest and digest bacteria thorough anaerobic pathways that process diverse energy sources. The bifunctional alcohol/aldehyde dehydrogenase enzyme in Entamoeba histolytica (EhADH2) belongs to the ADHE iron dependent family, and is essential for trophozoite growth and survival. EhADH2 catalyzes the conversion of acetyl Co‐A to acetaldehyde and the final reduction of acetaldehyde to ethanol by its separate ADH and ALDH domains respectively. Several Entamoeba spp have homologous genes (ADHE) which may have similar structure and function in the glycolytic pathway to that of E. histolytica . This study aims to clone, express, and sequence the genes that encode for ADHE in three Entamoeba spp . The sequencing identifies evolutionary relationships between the homologs. The transformation of ADHE genes into E. coli cells will be used for protein purification and kinetic assays for comparison with Entamoeba histolytica ADHE (EhADH2) previously studied by this lab. These kinetic assays will provide Km values for both substrates and information on chemical inhibitors of the ADHE proteins. Inhibitors of this enzyme have promising anti‐amoebic capabilities for drugs due to ADHE necessity during anaerobic respiration. Anaerobic pathogens have evolved adaptive metabolic enzymes that differ from vertebrates, and are ideal targets for novel compounds. We are interested in furthering our knowledge about the bifunctional alcohol/aldehyde dehydrogenase EhADH2 enzyme (required for E. histolytica growth and survival) and its adaptations to develop improved/non‐toxic strategies to manage amebiasis in humans.