Existence in animal tissues of adenosine triphosphate thiamin diphosphate phosphotransferase [EC 2.7.4.15]

An enzyme which catalyzes the synthesis of thiamin triphosphate from thiamin diphosphate (TDP), thiamindiphosphate kinase (ATP:thiamin diphosphate phosphotransferase) [EC 2.7.4.15], was detected in animal tissues. The enzyme was partially purified (150-fold) from the cytosol fraction of guinea pig brain. The enzyme reaction required free (not protein-bound) TDP, ATP, Mg2+, and a cofactor, which is a low molecular weight and heat-stable compound. The enzyme activity was optimal at pH 11 and at 25 degrees C. A stoichiometric transfer of 32P from [gamma-32P]ATP to TDP was demonstrated. Km values for TDP and ATP were calculated to be 1.1 mM and 10 microM, respectively, and Vmax was 868 nmol/mg of protein/hr. The enzyme was found solely in the cytosol fraction of guinea pig brain and was also detectable in the skeletal muscle and heart. These results provide strong evidence for the existence of TDP kinase in animal tissues.