Structure of the regulatory complex of Escherichia coli IIIGlc with glycerol kinase

The phosphocarrier protein IIIGlc is an integral component of the bacterial phosphotransferase (PTS) system. Unphosphorylated IIIGlc inhibits non-PTS carbohydrate transport systems by binding to diverse target proteins. The crystal structure at 2.6 A resolution of one of the targets, glycerol kinase (GK), in complex with unphosphorylated IIIGlc, glycerol, and adenosine diphosphate was determined. GK contains a region that is topologically identical to the adenosine triphosphate binding domains of hexokinase, the 70-kD heat shock cognate, and actin. IIIGlc binds far from the catalytic site of GK, indicating that long-range conformational changes mediate the inhibition of GK by IIIGlc. GK and IIIGlc are bound by hydrophobic and electrostatic interactions, with only one hydrogen bond involving an uncharged group. The phosphorylation site of IIIGlc, His90, is buried in a hydrophobic environment formed by the active site region of IIIGlc and a 3(10) helix of GK, suggesting that phosphorylation prevents IIIGlc binding to GK by directly disrupting protein-protein interactions.

[1]  W. Kabsch,et al.  Similarity of the three-dimensional structures of actin and the ATPase fragment of a 70-kDa heat shock cognate protein. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Michael G. Rossmann,et al.  Processing oscillation diffraction data for very large unit cells with an automatic convolution technique and profile fitting , 1979 .

[3]  R. St Charles,et al.  The three-dimensional structure of bovine platelet factor 4 at 3.0-A resolution. , 1989, The Journal of biological chemistry.

[4]  B. Lee,et al.  The interpretation of protein structures: estimation of static accessibility. , 1971, Journal of molecular biology.

[5]  A. Gronenborn,et al.  Solution structure of a calmodulin-target peptide complex by multidimensional NMR. , 1994, Science.

[6]  T. Steitz,et al.  A refined model of the sugar binding site of yeast hexokinase B. , 1978, Journal of molecular biology.

[7]  T. Steitz,et al.  Structure of a complex between yeast hexokinase A and glucose. I. Structure determination and refinement at 3.5 A resolution. , 1980, Journal of molecular biology.

[8]  S. Roseman,et al.  The bacterial phosphoenolpyruvate: glycose phosphotransferase system. , 1990, Annual review of biochemistry.

[9]  M. Karplus,et al.  Crystallographic R Factor Refinement by Molecular Dynamics , 1987, Science.

[10]  K. Flaherty,et al.  Three-dimensional structure of the ATPase fragment of a 70K heat-shock cognate protein , 1990, Nature.

[11]  C. Anfinsen,et al.  Limited proteolysis of IIIGlc, a regulatory protein of the phosphoenolpyruvate:glycose phosphotransferase system, by membrane-associated enzymes from Salmonella typhimurium and Escherichia coli. , 1986, The Journal of biological chemistry.

[12]  A M Gronenborn,et al.  Determination of the secondary structure of interleukin-8 by nuclear magnetic resonance spectroscopy. , 1989, The Journal of biological chemistry.

[13]  W. Kabsch,et al.  Atomic structure of the actin: DNase I complex , 1990, Nature.

[14]  S J Remington,et al.  Three-dimensional structure of the Escherichia coli phosphocarrier protein IIIglc. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[15]  Klevit Re,et al.  Two-dimensional 1H NMR studies of histidine-containing protein from Escherichia coli. 3. Secondary and tertiary structure as determined by NMR. , 1986 .

[16]  H. Paulus,et al.  Subunit dissociation in the allosteric regulation of glycerol kinase from Escherichia coli. 2. Physical evidence. , 1978, Biochemistry.

[17]  M H Saier,et al.  Structure of the IIA domain of the glucose permease of Bacillus subtilis at 2.2-A resolution. , 1991, Biochemistry.

[18]  D. Koshland,et al.  Electrostatic and steric contributions to regulation at the active site of isocitrate dehydrogenase. , 1990, Science.

[19]  D. Torchia,et al.  1H, 15N, and 13C NMR signal assignments of IIIGlc, a signal-transducing protein of Escherichia coli, using three-dimensional triple-resonance techniques. , 1991, Biochemistry.

[20]  R. Hamlin,et al.  [27] Multiwire area X-ray diffractometers , 1985 .

[21]  Lars Liljas,et al.  The three-dimensional structure of the bacterial virus MS2 , 1990, Nature.

[22]  B. Matthews,et al.  An oscillation data collection system for high‐resolution protein crystallography , 1981 .

[23]  D. Torchia,et al.  Secondary structure of the phosphocarrier protein IIIGlc, a signal-transducing protein from Escherichia coli, determined by heteronuclear three-dimensional NMR spectroscopy. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[24]  B. C. Wang.  Resolution of phase ambiguity in macromolecular crystallography. , 1985, Methods in enzymology.

[25]  C. Chothia,et al.  The structure of protein-protein recognition sites. , 1990, The Journal of biological chemistry.

[26]  M H Saier,et al.  Structure of the histidine-containing phosphocarrier protein HPr from Bacillus subtilis at 2.0-A resolution. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[27]  M. Karplus,et al.  Crystallographic refinement by simulated annealing: application to crambin , 1989 .

[28]  S. Remington,et al.  Crystallization and preliminary X-ray studies of Escherichia coli glycerol kinase. , 1989, Journal of molecular biology.

[29]  N. Xuong,et al.  Software for a diffractometer with multiwire area detector. , 1985, Methods in enzymology.

[30]  S. Roseman,et al.  Site-directed mutagenesis of the phosphocarrier protein. IIIGlc, a major signal-transducing protein in Escherichia coli. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[31]  M. Saier,et al.  Allosteric regulation of glycerol kinase by enzyme IIIglc of the phosphotransferase system in Escherichia coli and Salmonella typhimurium , 1985, Journal of bacteriology.

[32]  M. Saier,et al.  Functional interactions between proteins of the phosphoenolpyruvate:sugar phosphotransferase systems of Bacillus subtilis and Escherichia coli. , 1992, The Journal of biological chemistry.

[33]  S. Roseman,et al.  Signal transduction by the bacterial phosphotransferase system. Diauxie and the crr gene (J. Monod revisited). , 1990, The Journal of biological chemistry.

[34]  P. Postma,et al.  Interaction between IIIGlc of the phosphoenolpyruvate:sugar phosphotransferase system and glycerol kinase of Salmonella typhimurium , 1984, Journal of bacteriology.

[35]  D. Koshland,et al.  Regulation of isocitrate dehydrogenase by phosphorylation involves no long-range conformational change in the free enzyme. , 1991, The Journal of biological chemistry.

[36]  M. A. Saper,et al.  Structure of the human class I histocompatibility antigen, HLA-A2 , 1987, Nature.

[37]  D E Koshland,et al.  Regulation of an enzyme by phosphorylation at the active site. , 1991, Science.

[38]  P. Postma,et al.  Phosphoenolpyruvate:carbohydrate phosphotransferase system of bacteria. , 1985, Microbiological reviews.

[39]  J. Thorner,et al.  Catalytic and allosteric properties of glycerol kinase from Escherichia coli. , 1973, The Journal of biological chemistry.

[40]  D E Koshland,et al.  Structure of a bacterial enzyme regulated by phosphorylation, isocitrate dehydrogenase. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[41]  D. Torchia,et al.  Structural comparison of phosphorylated and unphosphorylated forms of IIIGlc, a signal-transducing protein from Escherichia coli, using three-dimensional NMR techniques. , 1992, Biochemistry.

[42]  David Eisenberg,et al.  Unbiased three-dimensional refinement of heavy-atom parameters by correlation of origin-removed Patterson functions , 1983 .

[43]  R. Huber,et al.  Crystal structure determination, refinement and molecular model of creatine amidinohydrolase from Pseudomonas putida. , 1988, Journal of molecular biology.

[44]  K. Entian,et al.  The primary structure of the yeast hexokinase PII gene (HXK2) which is responsible for glucose repression. , 1985, Gene.

[45]  H. Paulus,et al.  Subunit dissociation in the allosteric regulation of Glycerol kinase from Escherichia coli. 3. Role in desensitization. , 1978, Biochemistry.

[46]  Jones Ta,et al.  Diffraction methods for biological macromolecules. Interactive computer graphics: FRODO. , 1985, Methods in enzymology.

[47]  M. Saier.  Protein phosphorylation and allosteric control of inducer exclusion and catabolite repression by the bacterial phosphoenolpyruvate: sugar phosphotransferase system. , 1989, Microbiological reviews.