2au7: Difference between revisions
New page: left|200px<br /><applet load="2au7" size="450" color="white" frame="true" align="right" spinBox="true" caption="2au7, resolution 1.05Å" /> '''The R43Q active site... |
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[[Image:2au7.gif|left|200px]]<br /><applet load="2au7" size=" | [[Image:2au7.gif|left|200px]]<br /><applet load="2au7" size="350" color="white" frame="true" align="right" spinBox="true" | ||
caption="2au7, resolution 1.05Å" /> | caption="2au7, resolution 1.05Å" /> | ||
'''The R43Q active site variant of E.coli inorganic pyrophosphatase'''<br /> | '''The R43Q active site variant of E.coli inorganic pyrophosphatase'''<br /> | ||
==Overview== | |||
Here, we describe high-resolution X-ray structures of Escherichia coli, inorganic pyrophosphatase (E-PPase) complexed with the substrate, magnesium, or manganese pyrophosphate. The structures correspond to steps, in the catalytic synthesis of enzyme-bound pyrophosphate (PP(i)) in the, presence of fluoride as an inhibitor of hydrolysis. The catalytic reaction, intermediates were trapped applying a new method that we developed for, initiating hydrolytic activity in the E-PPase crystal. X-ray structures, were obtained for three consecutive states of the enzyme in the course of, hydrolysis. Comparative analysis of these structures showed that the, Mn2+-supported hydrolysis of the phosphoanhydride bond is followed by a, fast release of the leaving phosphate from the P1 site. The electrophilic, phosphate P2 is trapped in the "down" conformation. Its movement into the, "up" position most likely represents the rate-limiting step of, Mn2+-supported hydrolysis. We further determined the crystal structure of, the Arg43Gln mutant variant of E-PPase complexed with one phosphate and, four Mn ions. | |||
==About this Structure== | ==About this Structure== | ||
2AU7 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 PO4, MN, NA and CL as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Inorganic_diphosphatase Inorganic diphosphatase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.6.1.1 3.6.1.1] Full crystallographic information is available from [http:// | 2AU7 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=PO4:'>PO4</scene>, <scene name='pdbligand=MN:'>MN</scene>, <scene name='pdbligand=NA:'>NA</scene> and <scene name='pdbligand=CL:'>CL</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Inorganic_diphosphatase Inorganic diphosphatase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.6.1.1 3.6.1.1] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2AU7 OCA]. | ||
==Reference== | |||
Reversible inhibition of Escherichia coli inorganic pyrophosphatase by fluoride: trapped catalytic intermediates in cryo-crystallographic studies., Samygina VR, Moiseev VM, Rodina EV, Vorobyeva NN, Popov AN, Kurilova SA, Nazarova TI, Avaeva SM, Bartunik HD, J Mol Biol. 2007 Mar 2;366(4):1305-17. Epub 2006 Dec 2. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17196979 17196979] | |||
[[Category: Escherichia coli]] | [[Category: Escherichia coli]] | ||
[[Category: Inorganic diphosphatase]] | [[Category: Inorganic diphosphatase]] | ||
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[[Category: mutant]] | [[Category: mutant]] | ||
''Page seeded by [http:// | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Jan 23 11:17:26 2008'' | ||
