2rkd
From Proteopedia
(New page: 200px<br /><applet load="2rkd" size="350" color="white" frame="true" align="right" spinBox="true" caption="2rkd, resolution 1.900Å" /> '''The Structure of ra...) |
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==Overview== | ==Overview== | ||
| - | The mechanisms of molecular recognition of phosphoenolpyruvate (PEP) and | + | The mechanisms of molecular recognition of phosphoenolpyruvate (PEP) and oxaloacetate (OAA) by cytosolic phosphoenolpyruvate carboxykinase (cPEPCK) were investigated by the systematic evaluation of a variety of PEP and OAA analogues as potential reversible inhibitors of the enzyme against PEP. The molecules that inhibit the enzyme in a competitive fashion were found to fall into two general classes. Those molecules that mimic the binding geometry of PEP, namely phosphoglycolate and 3-phosphonopropionate, are found to bind weakly (millimolar Ki values). In contrast, those competitive inhibitors that mimic the binding of OAA (oxalate and phosphonoformate) coordinate directly to the active site manganese ion and bind an order of magnitude more tightly (micromolar Ki values). The competitive inhibitor sulfoacetate is found to be an outlier of these two classes, binding in a hybrid fashion utilizing modes of recognition of both PEP and OAA in order to achieve a micromolar inhibition constant in the absence of direct coordination to the active site metal. The kinetic studies in combination with the structural characterization of the five aforementioned competitive inhibitors demonstrate the molecular requirements for high affinity binding of molecules to the active site of the enzyme. These features include cis-planar carbonyl groups that are required for coordination to the active site metal, a bridging electron rich atom at the position corresponding to the C2 methylene group of OAA to facilitate interactions with R405, a carboxylate or sulfonate moiety at a position corresponding to the C1 carboxylate of OAA, and the edge-on aromatic interaction between a carboxylate and Y235. |
==About this Structure== | ==About this Structure== | ||
| - | 2RKD is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Rattus_norvegicus Rattus norvegicus] with <scene name='pdbligand=MN:'>MN</scene>, <scene name='pdbligand=NA:'>NA</scene> and <scene name='pdbligand=3PP:'>3PP</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Phosphoenolpyruvate_carboxykinase_(GTP) Phosphoenolpyruvate carboxykinase (GTP)], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=4.1.1.32 4.1.1.32] Known structural/functional Sites: <scene name='pdbsite=AC1:Mn Binding Site For Residue A 700'>AC1</scene>, <scene name='pdbsite=AC2:Mn Binding Site For Residue A 701'>AC2</scene>, <scene name='pdbsite=AC3:Na Binding Site For Residue A 702'>AC3</scene> and <scene name='pdbsite=AC4:3pp Binding Site For Residue A 703'>AC4</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2RKD OCA]. | + | 2RKD is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Rattus_norvegicus Rattus norvegicus] with <scene name='pdbligand=MN:'>MN</scene>, <scene name='pdbligand=NA:'>NA</scene> and <scene name='pdbligand=3PP:'>3PP</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Phosphoenolpyruvate_carboxykinase_(GTP) Phosphoenolpyruvate carboxykinase (GTP)], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=4.1.1.32 4.1.1.32] Known structural/functional Sites: <scene name='pdbsite=AC1:Mn+Binding+Site+For+Residue+A+700'>AC1</scene>, <scene name='pdbsite=AC2:Mn+Binding+Site+For+Residue+A+701'>AC2</scene>, <scene name='pdbsite=AC3:Na+Binding+Site+For+Residue+A+702'>AC3</scene> and <scene name='pdbsite=AC4:3pp+Binding+Site+For+Residue+A+703'>AC4</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2RKD OCA]. |
==Reference== | ==Reference== | ||
| - | Differential Inhibition of Cytosolic PEPCK by Substrate Analogues. Kinetic and Structural Characterization of Inhibitor Recognition., Stiffin RM, Sullivan SM, Carlson GM, Holyoak T, Biochemistry. 2008 Jan 16 | + | Differential Inhibition of Cytosolic PEPCK by Substrate Analogues. Kinetic and Structural Characterization of Inhibitor Recognition., Stiffin RM, Sullivan SM, Carlson GM, Holyoak T, Biochemistry. 2008 Feb 19;47(7):2099-109. Epub 2008 Jan 16. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=18197707 18197707] |
[[Category: Phosphoenolpyruvate carboxykinase (GTP)]] | [[Category: Phosphoenolpyruvate carboxykinase (GTP)]] | ||
[[Category: Rattus norvegicus]] | [[Category: Rattus norvegicus]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| - | [[Category: Carlson, G | + | [[Category: Carlson, G M.]] |
[[Category: Holyoak, T.]] | [[Category: Holyoak, T.]] | ||
| - | [[Category: Stiffin, R | + | [[Category: Stiffin, R M.]] |
| - | [[Category: Sullivan, S | + | [[Category: Sullivan, S M.]] |
[[Category: 3PP]] | [[Category: 3PP]] | ||
[[Category: MN]] | [[Category: MN]] | ||
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[[Category: nucleotide-binding]] | [[Category: nucleotide-binding]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:48:06 2008'' |
Revision as of 16:48, 21 February 2008
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The Structure of rat cytosolic PEPCK in complex with 3-phosphonopropionate
Overview
The mechanisms of molecular recognition of phosphoenolpyruvate (PEP) and oxaloacetate (OAA) by cytosolic phosphoenolpyruvate carboxykinase (cPEPCK) were investigated by the systematic evaluation of a variety of PEP and OAA analogues as potential reversible inhibitors of the enzyme against PEP. The molecules that inhibit the enzyme in a competitive fashion were found to fall into two general classes. Those molecules that mimic the binding geometry of PEP, namely phosphoglycolate and 3-phosphonopropionate, are found to bind weakly (millimolar Ki values). In contrast, those competitive inhibitors that mimic the binding of OAA (oxalate and phosphonoformate) coordinate directly to the active site manganese ion and bind an order of magnitude more tightly (micromolar Ki values). The competitive inhibitor sulfoacetate is found to be an outlier of these two classes, binding in a hybrid fashion utilizing modes of recognition of both PEP and OAA in order to achieve a micromolar inhibition constant in the absence of direct coordination to the active site metal. The kinetic studies in combination with the structural characterization of the five aforementioned competitive inhibitors demonstrate the molecular requirements for high affinity binding of molecules to the active site of the enzyme. These features include cis-planar carbonyl groups that are required for coordination to the active site metal, a bridging electron rich atom at the position corresponding to the C2 methylene group of OAA to facilitate interactions with R405, a carboxylate or sulfonate moiety at a position corresponding to the C1 carboxylate of OAA, and the edge-on aromatic interaction between a carboxylate and Y235.
About this Structure
2RKD is a Single protein structure of sequence from Rattus norvegicus with , and as ligands. Active as Phosphoenolpyruvate carboxykinase (GTP), with EC number 4.1.1.32 Known structural/functional Sites: , , and . Full crystallographic information is available from OCA.
Reference
Differential Inhibition of Cytosolic PEPCK by Substrate Analogues. Kinetic and Structural Characterization of Inhibitor Recognition., Stiffin RM, Sullivan SM, Carlson GM, Holyoak T, Biochemistry. 2008 Feb 19;47(7):2099-109. Epub 2008 Jan 16. PMID:18197707
Page seeded by OCA on Thu Feb 21 18:48:06 2008
