title: An alkali metal ion size-dependent switch in the active site structure of dialkylglycine decarboxylase.
abstract: The pyridoxal 5'-phosphate-dependent enzyme dialkylglycine decarboxylase (DGD) is activated by K+ and Rb+ ions, whereas Li+ and Na+ ions are inhibitory. A binding site for alkali metal ions close to the active site (site 1) was discovered in the crystal structure of DGD, and an exchange of K+ for Na+ at this site was shown to affect the conformation of two active site residues [Toney, M. D., Hohenester, E., Cowan, S. W., & Jansonius, J. N. (1993) Science 261, 756-759]. We have investigated the effects of alkali metal ions on DGD activity and have determined the crystal structures at 2.8 A resolution of DGD with Li+ and Rb+ bound at site 1. Due to the weak scattering of the Li+ ion, its position had to be modeled using information from small molecule structures. A comparison of the DGD structures with Li+, Na+, K+, and Rb+ bound at site 1 reveals a striking correlation between active site structure and enzymatic activity. The small, inhibitory ions Li+ and Na+ are accommodated by replacing two protein-derived ligands of the larger, activating ions K+ and Rb+ by a single water molecule. This actuates a two-state structural switch between active and inactive enzyme that involves a concerted reorientation of the active site residues Ser80 and Tyr301 and a small change in the quaternary structure of the DGD tetramer. An important role of the essential K+ ion in both cofactor binding and the organization of a catalytically competent active site structure is proposed. In the structure of DGD with Rb+ bound at site 1, a second Rb+ ion has partially replaced the structural Na+ ion at metal binding site 2 on the surface of the DGD molecule, without significantly altering the protein structure. In contrast to Na+, the Rb+ ion is bound with unfavorable geometry, and it is proposed that the rigid site 2 structure results in a pronounced selectivity for Na+ ions.
authors.affiliation: Department of Structural Biology, University of Basel, Switzerland.
authors.forename: E
authors.initials: E
authors.lastname: Hohenester
authors.forename: J W
authors.initials: JW
authors.lastname: Keller
authors.forename: J N
authors.initials: JN
authors.lastname: Jansonius
mesh_terms: D001665:Binding Sites; D002262:Carboxy-Lyases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D003196:Computer Graphics; D018360:Crystallography, X-Ray; D004789:Enzyme Activation; D007477:Ions; D008672:Metals, Alkali / Q000737:chemistry*; D011487:Protein Conformation
chemical_list: D007477:Ions; D008672:Metals, Alkali; D002262:Carboxy-Lyases; C063519:2,2-dialkylglycine decarboxylase