Half-of-the-site reactivity of the decarboxylating component of the pyruvate dehydrogenase complex from pigeon breast muscle with respect to 2-hydroxyethyl thiamine pyrophosphate.

The holopyruvate dehydrogenase is characterized by the charge transfer complex formation between tryptophan residue and thiamine pyrophosphate in each of two active centres. Interaction of apoenzyme with one mole of 2-hydroxyethyl thiamine pyrophosphate results in appearance of the same spectral band which does not change in intensity with further increase in ligand concentration. 2-hydroxyethyl thiamine pyrophosphate: acceptor oxidoreductase activity abolishes after oxidation of only one tryptophan residue per mole of the protein or blocking of one of the active centres with inactive analogue of the coenzyme. In the latter case the charge transfer complex band induced by interaction of apoenzyme with 2-hydroxyethyl thiamine pyrophosphate was not shown at all. These facts testify to half-of-the-site reactivity of pyruvate dehydrogenase with respect to 2-hydroxyethyl thiamine pyrophosphate.