Molecular and biochemical characterization of phosphoglucomutases from Entamoeba histolytica and Entamoeba dispar.
Entamoeba histolytica and Entamoeba dispar have only recently been defined as two separate species. E. histolytica, the pathogenic species, is the microorganism causing invasive intestinal amoebiasis and/or liver abscess, while the morphologically similar E. dispar is nonpathogenic and noninvasive. The gold standard for the distinction of the two species has been the isoenzyme electrophoresis of phosphoglucomutases (EC 5.4.2.2) and hexokinases (EC 2.7.1.1), but there had also been a controversy about the possibility of a conversion of isoenzyme patterns. In this study, we cloned the phosphoglucomutase (PGM) cDNAs from the pathogenic and the nonpathogenic species. The deduced amino acid sequences were only 2.4% different. The cDNAs were expressed in Escherichia coli under the control of a T7 RNA polymerase promoter. The recombinant polypeptides displayed strong phosphoglucomutase activity, each of the recombinant enzymes comigrated with its natural counterpart from E. histolytica and E. dispar in the starch gel electrophoresis. Our results give a biochemical interpretation of the PGM isoenzyme pattern and support the clear distinction between the two species.
Ortner
S
S
Institute for Specific Prophylaxis and Tropical Medicine, University of Vienna, Austria.
Binder
M
M
Scheiner
O
O
Wiedermann
G
G
Duchêne
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004592:Electrophoresis, Starch Gel; D004747:Entamoeba / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D004748:Entamoeba histolytica / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D017125:Genes, Protozoan; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010733:Phosphoglucomutase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D018076:DNA, Complementary; D007527:Isoenzymes; D011994:Recombinant Proteins; D010733:Phosphoglucomutase