1bo5: Difference between revisions

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CRYSTAL STRUCTURE OF THE COMPLEX BETWEEN ESCHERICHIA COLI GLYCEROL KINASE AND THE ALLOSTERIC REGULATOR FRUCTOSE 1,6-BISPHOSPHATE.

OverviewOverview

The three-dimensional structures of Escherichia coli glycerol kinase (GK) with bound glycerol in the presence and absence of one of the allosteric regulators of its activity, fructose 1,6-bisphosphate (FBP), at 3.2 and 3.0 A, are presented. The molecule crystallized in space group P41212, and the structure was solved by molecular replacement. The models were refined with good stereochemistry to final R-factors of 21.1 and 21.9%, respectively. A tetrameric arrangement of monomers was observed which was essentially identical to the proposed inactive tetramer II previously described [Feese, M. D., Faber, H. R., Bystrom, C. E., Pettigrew, D. W., and Remington, S. J. (1998) Structure (in press)]. However, the crystal packing in this form was especially open, permitting the FBP binding site to be occupied and identified. The crystallographic data revealed a most unusual type of FBP binding site formed between two glycine-arginine loops (residues 234-236) where one-half of the binding site is donated by each monomer at the regulatory interface. The molecule of FBP binds in two mutually exclusive modes on a noncrystallographic 2-fold axis at 50% occupancy each; thus, a tetramer of GK binds two molecules of FBP. Ionic interactions between the 1- and 6-phosphates of FBP and Arg 236 were observed in addition to hydrogen bonding interactions between the backbone amide of Gly 234 and the 6-phosphate. No contacts between the protein and the furanose ring were observed. Mutagenesis of Arg 236 to alanine drastically reduced the extent of inhibition of GK by FBP and lowered, but did not eliminate, the ability of FBP to promote tetramer association. These observations are consistent with the previously characterized mechanism of FBP inhibition of GK, in which FBP acts both to promote dimer-tetramer assembly and to inactivate the tetramers.

About this StructureAbout this Structure

1BO5 is a Single protein structure of sequence from Escherichia coli with and as ligands. Active as Glycerol kinase, with EC number 2.7.1.30 Full crystallographic information is available from OCA.

ReferenceReference

Crystal structure of a complex of Escherichia coli glycerol kinase and an allosteric effector fructose 1,6-bisphosphate., Ormo M, Bystrom CE, Remington SJ, Biochemistry. 1998 Nov 24;37(47):16565-72. PMID:9843423

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-[[Image:1bo5.jpg|left|200px]]<br /><applet load="1bo5" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:1bo5.jpg|left|200px]]<br /><applet load="1bo5" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="1bo5, resolution 3.2&Aring;" />
 '''CRYSTAL STRUCTURE OF THE COMPLEX BETWEEN ESCHERICHIA COLI GLYCEROL KINASE AND THE ALLOSTERIC REGULATOR FRUCTOSE 1,6-BISPHOSPHATE.'''<br />
 ==Overview==
-The three-dimensional structures of Escherichia coli glycerol kinase (GK), with bound glycerol in the presence and absence of one of the allosteric, regulators of its activity, fructose 1,6-bisphosphate (FBP), at 3.2 and, 3.0 A, are presented. The molecule crystallized in space group P41212, and, the structure was solved by molecular replacement. The models were refined, with good stereochemistry to final R-factors of 21.1 and 21.9%, respectively. A tetrameric arrangement of monomers was observed which was, essentially identical to the proposed inactive tetramer II previously, described [Feese, M. D., Faber, H. R., Bystrom, C. E., Pettigrew, D. W., and Remington, S. J. (1998) Structure (in press)]. However, the crystal, packing in this form was especially open, permitting the FBP binding site, to be occupied and identified. The crystallographic data revealed a most, unusual type of FBP binding site formed between two glycine-arginine loops, (residues 234-236) where one-half of the binding site is donated by each, monomer at the regulatory interface. The molecule of FBP binds in two, mutually exclusive modes on a noncrystallographic 2-fold axis at 50%, occupancy each; thus, a tetramer of GK binds two molecules of FBP. Ionic, interactions between the 1- and 6-phosphates of FBP and Arg 236 were, observed in addition to hydrogen bonding interactions between the backbone, amide of Gly 234 and the 6-phosphate. No contacts between the protein and, the furanose ring were observed. Mutagenesis of Arg 236 to alanine, drastically reduced the extent of inhibition of GK by FBP and lowered, but, did not eliminate, the ability of FBP to promote tetramer association., These observations are consistent with the previously characterized, mechanism of FBP inhibition of GK, in which FBP acts both to promote, dimer-tetramer assembly and to inactivate the tetramers.
+The three-dimensional structures of Escherichia coli glycerol kinase (GK) with bound glycerol in the presence and absence of one of the allosteric regulators of its activity, fructose 1,6-bisphosphate (FBP), at 3.2 and 3.0 A, are presented. The molecule crystallized in space group P41212, and the structure was solved by molecular replacement. The models were refined with good stereochemistry to final R-factors of 21.1 and 21.9%, respectively. A tetrameric arrangement of monomers was observed which was essentially identical to the proposed inactive tetramer II previously described [Feese, M. D., Faber, H. R., Bystrom, C. E., Pettigrew, D. W., and Remington, S. J. (1998) Structure (in press)]. However, the crystal packing in this form was especially open, permitting the FBP binding site to be occupied and identified. The crystallographic data revealed a most unusual type of FBP binding site formed between two glycine-arginine loops (residues 234-236) where one-half of the binding site is donated by each monomer at the regulatory interface. The molecule of FBP binds in two mutually exclusive modes on a noncrystallographic 2-fold axis at 50% occupancy each; thus, a tetramer of GK binds two molecules of FBP. Ionic interactions between the 1- and 6-phosphates of FBP and Arg 236 were observed in addition to hydrogen bonding interactions between the backbone amide of Gly 234 and the 6-phosphate. No contacts between the protein and the furanose ring were observed. Mutagenesis of Arg 236 to alanine drastically reduced the extent of inhibition of GK by FBP and lowered, but did not eliminate, the ability of FBP to promote tetramer association. These observations are consistent with the previously characterized mechanism of FBP inhibition of GK, in which FBP acts both to promote dimer-tetramer assembly and to inactivate the tetramers.
 ==About this Structure==
-BO5 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli] with FBP and GOL as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Glycerol_kinase Glycerol kinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.1.30 2.7.1.30] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1BO5 OCA].
+BO5 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli] with <scene name='pdbligand=FBP:'>FBP</scene> and <scene name='pdbligand=GOL:'>GOL</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Glycerol_kinase Glycerol kinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.1.30 2.7.1.30] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1BO5 OCA].
 ==Reference==
@@ Line 14: / Line 14: @@
 [[Category: Glycerol kinase]]
 [[Category: Single protein]]
-[[Category: Bystrom, C.E.]]
+[[Category: Bystrom, C E.]]
 [[Category: Ormo, M.]]
-[[Category: Remington, S.J.]]
+[[Category: Remington, S J.]]
 [[Category: FBP]]
 [[Category: GOL]]
@@ Line 23: / Line 23: @@
 [[Category: kinase]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Nov 20 11:47:07 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 11:57:19 2008''