A THIAMIN DIPHOSPHATE BINDING FOLD REVEALED BY COMPARISON OF THE CRYSTAL-STRUCTURES OF TRANSKETOLASE, PYRUVATE OXIDASE AND PYRUVATE DECARBOXYLASE

Background: The crystal structures of three thiamin diphosphate-dependent enzymes that catalyze distinct reactions in basic metabolic pathways are known. These enzymes - transketolase, pyruvate oxidase and pyruvate decarboxylase - also require metal ions such as Ca2+ and Mg2+ as cofactors and have little overall sequence similarity. Here, the crystal structures of these three enzymes are compared. Results: The three enzymes share a similar pattern of binding of thiamin diphosphate and the metal ion cofactors. The enzymes function as multisubunit proteins, with each polypeptide chain folded into three alpha/beta domains. Two of these domains are involved in binding of the thiamin diphosphate and the metal ion. These domains have the same topology of six parallel beta-strands and surrounding alpha-helices. The thiamin diphosphate is bound in a cleft, formed by mio domains from two different subunits. Only a few residues are conserved in all three enzymes and these are responsible for proper binding of the cofactors. Conclusions: Despite considerable differences in qua ternary structure and lack of overall sequence homology, thiamin diphosphate binds to the three enzymes in a very similar fashion, and a general thiamin-binding fold can be revealed.