Modification of thiamine pyrophosphate dependent enzyme activity by oxythiamine in Saccharomyces cerevisiae cells.
Oxythiamine is an antivitamin derivative of thiamine that after phosphorylation to oxythiamine pyro phosphate can bind to the active centres of thiamine-dependent enzymes. In the present study, the effect of oxythiamine on the viability of Saccharomyces cerevisiae and the activity of thiamine pyrophosphate dependent enzymes in yeast cells has been investigated. We observed a decrease in pyruvate decarboxylase specific activity on both a control and an oxythiamine medium after the first 6 h of culture. The cytosolic enzymes transketolase and pyruvate decarboxylase decreased their specific activity in the presence of oxythiamine but only during the beginning of the cultivation. However, after 12 h of cultivation, oxythiamine-treated cells showed higher specific activity of cytosolic enzymes. More over, it was established by SDS-PAGE that the high specific activity of pyruvate decarboxylase was followed by an increase in the amount of the enzyme protein. In contrast, the mitochondrial enzymes, pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase complexes, were inhibited by oxythiamine during the entire experiment. Our results suggest that the observed strong decrease in growth rate and viability of yeast on medium with oxythiamine may be due to stronger inhibition of mitochondrial pyruvate dehydrogenase than of cytosolic enzymes.
Tylicki
Adam
A
Institute of Biology, University of Bialystok, Bialystok, Poland. atyl@uwb.edu.pl
Czerniecki
Jan
J
Dobrzyn
Pawel
P
Matanowska
Agnieszka
A
Olechno
Anna
A
Strumilo
Slawomir
S
D000963:Antimetabolites / Q000494:pharmacology*; D015169:Colony Count, Microbial; D003470:Culture Media; D003600:Cytosol / Q000201:enzymology; D007655:Ketoglutarate Dehydrogenase Complex / Q000187:drug effects / Q000378:metabolism; D008928:Mitochondria / Q000201:enzymology; D010119:Oxythiamine / Q000494:pharmacology*; D011767:Pyruvate Decarboxylase / Q000187:drug effects / Q000378:metabolism; D011768:Pyruvate Dehydrogenase Complex / Q000187:drug effects / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development*; D013835:Thiamine Pyrophosphate / Q000378:metabolism*; D014174:Transketolase / Q000187:drug effects / Q000378:metabolism
D000963:Antimetabolites; D003470:Culture Media; D011768:Pyruvate Dehydrogenase Complex; D010119:Oxythiamine; D007655:Ketoglutarate Dehydrogenase Complex; D014174:Transketolase; D011767:Pyruvate Decarboxylase; D013835:Thiamine Pyrophosphate