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10.1074/jbc.M115.636936 http://scihub22266oqcxt.onion/10.1074/jbc.M115.636936 C4400351!4400351 !25694425     free     free     free Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 243.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 243.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 243.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Warning: imagejpeg(C:\Inetpub\vhosts\kidney.de\httpdocs\phplern\25694425 .jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117       J+Biol+Chem 2015 ; 290 (16 ): 10418-29 Nephropedia Template TP {{tp|p=25694425 |t=2015. Structure of Escherichia coli dGTP triphosphohydrolase: a hexameric enzyme with DNA effector molecules |pdf=|usr=}}{{25694425 }} gab.com Text Structure of Escherichia coli dGTP triphosphohydrolase: a hexameric enzyme with DNA effector molecules JO: J Biol Chem http://www.kidney.de/pget.php?&tw=1&pmid=25694425 #FOAvip The Escherichia coli dgt gene encodes a dGTP triphosphohydrolase whose detailed role still remains to be determined. Deletion of dgt creates a mutator phenotype, indicating that the dGTPase has a fidelity role, possibly by affecting the cellular dNTP pool. In the present study, we have investigated the structure of the Dgt protein at 3.1-Å resolution. One of the obtained structures revealed a protein hexamer that contained two molecules of single-stranded DNA. The presence of DNA caused significant conformational changes in the enzyme, including in the catalytic site of the enzyme. Dgt preparations lacking DNA were able to bind single-stranded DNA with high affinity (Kd ? 50 nM). DNA binding positively affected the activity of the enzyme: dGTPase activity displayed sigmoidal (cooperative) behavior without DNA but hyperbolic (Michaelis-Menten) kinetics in its presence, consistent with a specific lowering of the apparent Km for dGTP. A mutant Dgt enzyme was also created containing residue changes in the DNA binding cleft. This mutant enzyme, whereas still active, was incapable of DNA binding and could no longer be stimulated by addition of DNA. We also created an E. coli strain containing the mutant dgt gene on the chromosome replacing the wild-type gene. The mutant also displayed a mutator phenotype. Our results provide insight into the allosteric regulation of the enzyme and support a physiologically important role of DNA binding. Twit Text FOAVip Structure of Escherichia coli dGTP triphosphohydrolase: a hexameric enzyme with DNA effector molecules ????J Biol Chem http://www.kidney.de/pget.php?&tw=1&pmid=25694425 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=25694425 #FOAvip English Wikipedia <ref>{{Cite pmid|25694425 }}</ref> Structure of Escherichia coli dGTP triphosphohydrolase: a hexameric enzyme with DNA effector molecules #MMPMID25694425 Singh D ; Gawel D ; Itsko M ; Hochkoeppler A ; Krahn JM ; London RE ; Schaaper RM J Biol Chem 2015[Apr]; 290 (16 ): 10418-29 PMID25694425 show ga The Escherichia coli dgt gene encodes a dGTP triphosphohydrolase whose detailed role still remains to be determined. Deletion of dgt creates a mutator phenotype, indicating that the dGTPase has a fidelity role, possibly by affecting the cellular dNTP pool. In the present study, we have investigated the structure of the Dgt protein at 3.1-Å resolution. One of the obtained structures revealed a protein hexamer that contained two molecules of single-stranded DNA. The presence of DNA caused significant conformational changes in the enzyme, including in the catalytic site of the enzyme. Dgt preparations lacking DNA were able to bind single-stranded DNA with high affinity (Kd ? 50 nM). DNA binding positively affected the activity of the enzyme: dGTPase activity displayed sigmoidal (cooperative) behavior without DNA but hyperbolic (Michaelis-Menten) kinetics in its presence, consistent with a specific lowering of the apparent Km for dGTP. A mutant Dgt enzyme was also created containing residue changes in the DNA binding cleft. This mutant enzyme, whereas still active, was incapable of DNA binding and could no longer be stimulated by addition of DNA. We also created an E. coli strain containing the mutant dgt gene on the chromosome replacing the wild-type gene. The mutant also displayed a mutator phenotype. Our results provide insight into the allosteric regulation of the enzyme and support a physiologically important role of DNA binding. |*Gene Expression Regulation, Bacterial [MESH] |Allosteric Regulation [MESH] |Catalytic Domain [MESH] |Chromosomes, Bacterial/chemistry/metabolism [MESH] |Crystallography, X-Ray [MESH] |DNA, Bacterial/*chemistry/metabolism [MESH] |DNA-Binding Proteins/chemistry/genetics/metabolism [MESH] |Deoxyguanine Nucleotides/*chemistry/metabolism [MESH] |Escherichia coli Proteins/*chemistry/genetics/metabolism [MESH] |Escherichia coli/*enzymology/genetics [MESH] |Kinetics [MESH] |Models, Molecular [MESH] |Mutation [MESH] |Phosphoric Monoester Hydrolases/*chemistry/genetics/metabolism [MESH] |Protein Multimerization [MESH] |Protein Structure, Secondary [MESH]Structure of Escherichia coli dGTP triphosphohydrolase: a hexameric en(epmc).pdf DeepDyve Pubget Overpricing