title: Molecular analysis of the cytosolic and glycosomal glyceraldehyde-3-phosphate dehydrogenase in Leishmania mexicana.
abstract: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity was detected in two cell compartments of Leishmania mexicana promastigotes. These activities could be attributed to two different isoenzymes, one residing in glycosomes, the other in the cytosol. We have cloned and sequenced the genes for both isoenzymes. The glycosomal enzyme is encoded by two tandemly linked genes of identical sequence and contains features frequently found in glycosomal enzymes: the presence of peptide insertions, a small carboxy-terminal extension with a potential glycosomal targeting signal (-SKM) and an excess of positively charged residues (net charge +7). Only one open reading frame was detected for the cytosolic enzyme. The amino acid sequences of the two proteins are only 55% identical. We discuss some evolutionary aspects of the observed organization of the GAPDH genes in the Trypanosomatidae and the role of the two isoenzymes in the metabolism of these organisms. The possibility to develop GAPDH-specific inhibitors that will be effective against the enzyme of various parasitic members of this family is explored.
authors.affiliation: International Institute of Cellular and Molecular Pathology, Research Unit for Tropical Diseases, Brussels, Belgium.
authors.forename: V
authors.initials: V
authors.lastname: Hannaert
authors.forename: M
authors.initials: M
authors.lastname: Blaauw
authors.forename: L
authors.initials: L
authors.lastname: Kohl
authors.forename: S
authors.initials: S
authors.lastname: Allert
authors.forename: F R
authors.initials: FR
authors.lastname: Opperdoes
authors.forename: P A
authors.initials: PA
authors.lastname: Michels
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D003600:Cytosol / Q000201:enzymology; D016054:DNA, Protozoan / Q000737:chemistry*; D004072:Digitonin; D004789:Enzyme Activation; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006593:Hexokinase / Q000378:metabolism; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics*; D007894:Leishmania mexicana / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D015388:Organelles / Q000201:enzymology; D010802:Phylogeny; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA
chemical_list: D003062:Codon; D016054:DNA, Protozoan; D007527:Isoenzymes; D005954:Glucosephosphate Dehydrogenase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D006593:Hexokinase; D004072:Digitonin