Structural basis for the ADP-specificity of a novel glucokinase from a hyperthermophilic archaeon

Background: ATP is the most common phosphoryl group donor for kinases. Howe ver, certain hyperthermophilic archaea such as Thermococcus litoralis and P yrococcus furiosus utilize unusual ADP-dependent glucokinases and phosphofr uctokinases in their glycolytic pathways. These ADP-dependent kinases are h omologous to each other but show no sequence similarity to any of the hithe rto known ATP-dependent enzymes. Results: We solved the crystal structure at 2.3 Angstrom resolution of an A DP-dependent glucokinase from T, litoralis (tIGK) complexed with ADP. The o verall structure can be divided into large and small alpha/beta domains, an d the ADP molecule is buried in a shallow pocket in the large domain. Unexp ectedly, the structure was similar to those of two ATP-dependent kinases, r ibokinase and adenosine kinase. Comparison based on three-dimensional struc ture revealed that several motifs important both in structure and function are conserved, and the recognition of the alpha- and beta -phosphate of the ADP in the tIGK was almost identical with the recognition of the beta- and gamma -phosphate of ATP in these ATP-dependent kinases. Conclusions: Noticeable points of our study are the first structure of ADP- dependent kinase, the structural similarity to members of the ATP-dependent ribokinase family, its rare nucleotide specificity caused by a shift in nu cleotide binding position by one phosphate unit, and identification of the residues that discriminate ADP-and ATP-dependence. The strict conservation of the binding site for the terminal and adjacent phosphate moieties sugges ts a common ancestral origin of both the ATP- and ADP-dependent kinases.