title: Triose phosphate isomerase deficiency is caused by altered dimerization--not catalytic inactivity--of the mutant enzymes.
abstract: Triosephosphate isomerase (TPI) deficiency is an autosomal recessive disorder caused by various mutations in the gene encoding the key glycolytic enzyme TPI. A drastic decrease in TPI activity and an increased level of its substrate, dihydroxyacetone phosphate, have been measured in unpurified cell extracts of affected individuals. These observations allowed concluding that the different mutations in the TPI alleles result in catalytically inactive enzymes. However, despite a high occurrence of TPI null alleles within several human populations, the frequency of this disorder is exceptionally rare. In order to address this apparent discrepancy, we generated a yeast model allowing us to perform comparative in vivo analyses of the enzymatic and functional properties of the different enzyme variants. We discovered that the majority of these variants exhibit no reduced catalytic activity per se. Instead, we observed, the dimerization behavior of TPI is influenced by the particular mutations investigated, and by the use of a potential alternative translation initiation site in the TPI gene. Additionally, we demonstrated that the overexpression of the most frequent TPI variant, Glu104Asp, which displays altered dimerization features, results in diminished endogenous TPI levels in mammalian cells. Thus, our results reveal that enzyme deregulation attributable to aberrant dimerization of TPI, rather than direct catalytic inactivation of the enzyme, underlies the pathogenesis of TPI deficiency. Finally, we discovered that yeast cells expressing a TPI variant exhibiting reduced catalytic activity are more resistant against oxidative stress caused by the thiol-oxidizing reagent diamide. This observed advantage might serve to explain the high allelic frequency of TPI null alleles detected among human populations.
authors.affiliation: Max Planck Institute for Molecular Genetics, Berlin, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Ralser
authors.forename: Gino
authors.initials: G
authors.lastname: Heeren
authors.forename: Michael
authors.initials: M
authors.lastname: Breitenbach
authors.forename: Hans
authors.initials: H
authors.lastname: Lehrach
authors.forename: Sylvia
authors.initials: S
authors.lastname: Krobitsch
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002522:Chlorocebus aethiops; D003958:Diamide / Q000494:pharmacology; D004351:Drug Resistance / Q000235:genetics; D005816:Genetic Complementation Test; D006801:Humans; D008958:Models, Molecular; D009154:Mutation; D020125:Mutation, Missense; D010442:Peptide Chain Initiation, Translational; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D014305:Triose-Phosphate Isomerase / Q000737:chemistry / Q000172:deficiency* / Q000235:genetics*; D020798:Two-Hybrid System Techniques
chemical_list: D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D013439:Sulfhydryl Reagents; D003958:Diamide; D014305:Triose-Phosphate Isomerase