title: Structures of enterococcal glycerol kinase in the absence and presence of glycerol: correlation of conformation to substrate binding and a mechanism of activation by phosphorylation.
abstract: The first structure of a glycerol kinase from a Gram-positive organism, Enterococcus casseliflavus, has been determined to 2.8 A resolution in the presence of glycerol and to 2.5 A resolution in the absence of substrate. The substrate-induced closure of 7 degrees is significantly smaller than that reported for hexokinase, a model for substrate-mediated domain closure that has been proposed for glycerol kinase. Despite the 78% level of sequence identity and conformational similarity in the catalytic cleft regions of the En. casseliflavus and Escherichia coli glycerol kinases, remarkable structural differences have now been identified. These differences correlate well with their divergent regulatory schemes of activation by phosphorylation in En. casseliflavus and allosteric inhibition in E. coli. On the basis of our structural results, we propose a mechanism by which the phosphorylation of a histidyl residue located 25 A from the active site results in a 10-15-fold increase in the activity of the enterococcal glycerol kinase.
authors.affiliation: Department of Molecular Biology, Cell Biology and Biochemistry, Brown University, Providence, Rhode Island 02912, USA. Joanne_Yeh@brown.edu
authors.forename: Joanne I
authors.initials: JI
authors.lastname: Yeh
authors.forename: Véronique
authors.initials: V
authors.lastname: Charrier
authors.forename: Joao
authors.initials: J
authors.lastname: Paulo
authors.forename: Lihui
authors.initials: L
authors.lastname: Hou
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Darbon
authors.forename: Al
authors.initials: A
authors.lastname: Claiborne
authors.forename: Wim G J
authors.initials: WG
authors.lastname: Hol
authors.forename: Josef
authors.initials: J
authors.lastname: Deutscher
mesh_terms: D000595:Amino Acid Sequence; D001051:Apoenzymes / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D019281:Dimerization; D016983:Enterococcus / Q000201:enzymology*; D004789:Enzyme Activation; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism; D005990:Glycerol / Q000737:chemistry* / Q000378:metabolism*; D005991:Glycerol Kinase / Q000737:chemistry* / Q000378:metabolism*; D008969:Molecular Sequence Data; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D015398:Signal Transduction; D013379:Substrate Specificity
chemical_list: D001051:Apoenzymes; D029968:Escherichia coli Proteins; C032242:crr protein, E coli; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; D005991:Glycerol Kinase; D005990:Glycerol