2a2k: Difference between revisions

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[[Image:2a2k.gif|left|200px]]<br />

[[Image:2a2k.gif|left|200px]]<br /><applet load="2a2k" size="350" color="white" frame="true" align="right" spinBox="true"

<applet load="2a2k" size="~~450~~" color="white" frame="true" align="right" spinBox="true"

caption="2a2k, resolution 1.52Å" />

'''Crystal Structure of an active site mutant, C473S, of Cdc25B Phosphatase Catalytic Domain'''<br />

==Overview==

Cdc25 phosphatases are key activators of the eukaryotic cell cycle and, compelling anticancer targets because their overexpression has been, associated with numerous cancers. However, drug discovery targeting these, phosphatases has been hampered by the lack of structural information about, how Cdc25s interact with their native protein substrates, the, cyclin-dependent kinases. Herein, we predict a docked orientation for, Cdc25B with its Cdk2-pTpY-CycA protein substrate by a rigid-body docking, method and refine the docked models with full-scale molecular dynamics, simulations and minimization. We validate the stable ensemble structure, experimentally by a variety of in vitro and in vivo techniques., Specifically, we compare our model with a crystal structure of the, substrate-trapping mutant of Cdc25B. We identify and validate in vivo a, novel hot-spot residue on Cdc25B (Arg492) that plays a central role in, protein substrate recognition. We identify a hot-spot residue on the, substrate Cdk2 (Asp206) and confirm its interaction with hot-spot residues, on Cdc25 using hot-spot swapping and double mutant cycles to derive, interaction energies. Our experimentally validated model is consistent, with previous studies of Cdk2 and its interaction partners and initiates, the opportunity for drug discovery of inhibitors that target the remote, binding sites of this protein-protein interaction.

Cdc25 phosphatases are key activators of the eukaryotic cell cycle and compelling anticancer targets because their overexpression has been associated with numerous cancers. However, drug discovery targeting these phosphatases has been hampered by the lack of structural information about how Cdc25s interact with their native protein substrates, the cyclin-dependent kinases. Herein, we predict a docked orientation for Cdc25B with its Cdk2-pTpY-CycA protein substrate by a rigid-body docking method and refine the docked models with full-scale molecular dynamics simulations and minimization. We validate the stable ensemble structure experimentally by a variety of in vitro and in vivo techniques. Specifically, we compare our model with a crystal structure of the substrate-trapping mutant of Cdc25B. We identify and validate in vivo a novel hot-spot residue on Cdc25B (Arg492) that plays a central role in protein substrate recognition. We identify a hot-spot residue on the substrate Cdk2 (Asp206) and confirm its interaction with hot-spot residues on Cdc25 using hot-spot swapping and double mutant cycles to derive interaction energies. Our experimentally validated model is consistent with previous studies of Cdk2 and its interaction partners and initiates the opportunity for drug discovery of inhibitors that target the remote binding sites of this protein-protein interaction.

==About this Structure==

2A2K is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with CL and SO4 as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Protein-tyrosine-phosphatase Protein-tyrosine-phosphatase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.3.48 3.1.3.48] Full crystallographic information is available from [http://~~ispc~~.weizmann.ac.il/oca-bin/ocashort?id=2A2K OCA].

2A2K is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=CL:'>CL</scene> and <scene name='pdbligand=SO4:'>SO4</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Protein-tyrosine-phosphatase Protein-tyrosine-phosphatase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.3.48 3.1.3.48] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2A2K OCA].

==Reference==

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[[Category: substrate trapping]]

''Page seeded by [http://~~ispc~~.weizmann.ac.il/oca OCA ] on ~~Mon Nov 12 20~~:45:~~15 2007~~''

''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 16:22:58 2008''

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@@ Line 1: / Line 1: @@
-[[Image:2a2k.gif|left|200px]]<br />
+[[Image:2a2k.gif|left|200px]]<br /><applet load="2a2k" size="350" color="white" frame="true" align="right" spinBox="true"
-<applet load="2a2k" size="450" color="white" frame="true" align="right" spinBox="true"
 caption="2a2k, resolution 1.52&Aring;" />
 '''Crystal Structure of an active site mutant, C473S, of Cdc25B Phosphatase Catalytic Domain'''<br />
 ==Overview==
-Cdc25 phosphatases are key activators of the eukaryotic cell cycle and, compelling anticancer targets because their overexpression has been, associated with numerous cancers. However, drug discovery targeting these, phosphatases has been hampered by the lack of structural information about, how Cdc25s interact with their native protein substrates, the, cyclin-dependent kinases. Herein, we predict a docked orientation for, Cdc25B with its Cdk2-pTpY-CycA protein substrate by a rigid-body docking, method and refine the docked models with full-scale molecular dynamics, simulations and minimization. We validate the stable ensemble structure, experimentally by a variety of in vitro and in vivo techniques., Specifically, we compare our model with a crystal structure of the, substrate-trapping mutant of Cdc25B. We identify and validate in vivo a, novel hot-spot residue on Cdc25B (Arg492) that plays a central role in, protein substrate recognition. We identify a hot-spot residue on the, substrate Cdk2 (Asp206) and confirm its interaction with hot-spot residues, on Cdc25 using hot-spot swapping and double mutant cycles to derive, interaction energies. Our experimentally validated model is consistent, with previous studies of Cdk2 and its interaction partners and initiates, the opportunity for drug discovery of inhibitors that target the remote, binding sites of this protein-protein interaction.
+Cdc25 phosphatases are key activators of the eukaryotic cell cycle and compelling anticancer targets because their overexpression has been associated with numerous cancers. However, drug discovery targeting these phosphatases has been hampered by the lack of structural information about how Cdc25s interact with their native protein substrates, the cyclin-dependent kinases. Herein, we predict a docked orientation for Cdc25B with its Cdk2-pTpY-CycA protein substrate by a rigid-body docking method and refine the docked models with full-scale molecular dynamics simulations and minimization. We validate the stable ensemble structure experimentally by a variety of in vitro and in vivo techniques. Specifically, we compare our model with a crystal structure of the substrate-trapping mutant of Cdc25B. We identify and validate in vivo a novel hot-spot residue on Cdc25B (Arg492) that plays a central role in protein substrate recognition. We identify a hot-spot residue on the substrate Cdk2 (Asp206) and confirm its interaction with hot-spot residues on Cdc25 using hot-spot swapping and double mutant cycles to derive interaction energies. Our experimentally validated model is consistent with previous studies of Cdk2 and its interaction partners and initiates the opportunity for drug discovery of inhibitors that target the remote binding sites of this protein-protein interaction.
 ==About this Structure==
-A2K is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with CL and SO4 as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Protein-tyrosine-phosphatase Protein-tyrosine-phosphatase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.3.48 3.1.3.48] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=2A2K OCA].
+A2K is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=CL:'>CL</scene> and <scene name='pdbligand=SO4:'>SO4</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Protein-tyrosine-phosphatase Protein-tyrosine-phosphatase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.3.48 3.1.3.48] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2A2K OCA].
 ==Reference==
@@ Line 31: / Line 30: @@
 [[Category: substrate trapping]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov 12 20:45:15 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 16:22:58 2008''