by hanging-drop-based sparse-matrix screening strategy. NAD kinase crystallized in complex with its substrate NAD, its product NADP, or two synthesized NAD mimics, at near 2 A resolution. D45N mutant using the I222 5'-thioacetyladenosine-bound crystal form, to 2.2 A resolution, and mutant H223E
apo form of Ppnk at 2.8 A resolution. Crystals belong to space group C2 (a=140.2, b=69.4, c=106.4 A, and beta=130°). Holo form of Ppnk complexed with NAD+ at 2.6 A resolution. Crystals belong space group to P62 (a=b=110.5 and c=108.9 A). Overall structures of apo-Ppnk and Ppnk-NAD consist of an N-domain (residues 1-138 and 279-283), a C-domain (residues 139-278), and a C-terminal tail (residues 284-307)
crystals of SeMet-containing PPNK_THEMA are obtained at room temperature in hanging drops. Crystal structure of inorganic polyphosphate/ATP-NAD kinase is determined at 2.3 A resolution. The crystal structure is solved using single-wavelength anomalous diffraction data collected at the Se absorption-peak wavelength in a state in which no cofactors or substrates are bound. It revealed that the 258-amino-acid protein is folded into two distinct domains. The N-terminal alpha/beta-domain spans the first 100 amino acids and the last 30 amino acids of the polypeptide and has several topological matches in the PDB, whereas the other domain, which spans the middle 130 residues, adopts a unique beta-sandwich architecture and only appreciably matches the recently deposited PDB structures of NAD kinases