Abstract
Thiamin pyrophosphokinase was purified approximately 120,000-fold from crude extracts of pig brain by acid and heat treatment, ammonium sulfate fractionation, DEAE-cellulose column chromatography and affinity chromatography using TMP-agarose. The purified enzyme was homogenous on disc gel electrophoresis and the molecular weight of the enzyme was estimated to be 50,000 by gel filtration with Sephadex G-100,whereas 30,500 by sodium dodecyl sulfate polyacrylamide gel electrophoresis. These results suggest that the pig brain thiamin pyrophosphokinase may consist of two identical polypeptide chains.
