1mc3

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(New page: 200px<br /><applet load="1mc3" size="450" color="white" frame="true" align="right" spinBox="true" caption="1mc3, resolution 2.6&Aring;" /> '''CRYSTAL STRUCTURE OF ...)
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[[Image:1mc3.jpg|left|200px]]<br /><applet load="1mc3" size="450" color="white" frame="true" align="right" spinBox="true"
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[[Image:1mc3.jpg|left|200px]]<br /><applet load="1mc3" size="350" color="white" frame="true" align="right" spinBox="true"
caption="1mc3, resolution 2.6&Aring;" />
caption="1mc3, resolution 2.6&Aring;" />
'''CRYSTAL STRUCTURE OF RFFH'''<br />
'''CRYSTAL STRUCTURE OF RFFH'''<br />
==Overview==
==Overview==
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The enzyme glucose-1-phosphate thymidylyltransferase (RffH), the product, of the rffh gene, catalyzes one of the steps in the synthesis of, enterobacterial common antigen (ECA), a cell surface glycolipid found in, Gram-negative enteric bacteria. In Escherichia coli two gene products, RffH and RmlA, catalyze the same enzymatic reaction and are homologous in, sequence; however, they are part of different operons and function in, different pathways. We report the crystal structure of RffH bound to, deoxythymidine triphosphate (dTTP), the phosphate donor, and Mg(2+), refined at 2.6 A to an R-factor of 22.3% (R(free) = 28.4%). The crystal, structure of RffH shows a tetrameric enzyme best described as a dimer of, dimers. Each monomer has an overall alpha/beta fold and consists of two, domains, a larger nucleotide binding domain (residues 1-115, 222-291) and, a smaller sugar-binding domain (116-221), with the active site located at, the domain interface. The Mg(2+) ion is coordinated by two conserved, aspartates and the alpha-phosphate of deoxythymidine triphosphate. Its, location corresponds well to that in a structurally similar domain of, N-acetylglucosamine-1-phosphate uridylyltransferase (GlmU). Analysis of, the RffH, RmlA, and GlmU complexes with substrates and products provides, an explanation for their different affinities for Mg(2+) and leads to a, proposal for the dynamics along the reaction pathway.
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The enzyme glucose-1-phosphate thymidylyltransferase (RffH), the product of the rffh gene, catalyzes one of the steps in the synthesis of enterobacterial common antigen (ECA), a cell surface glycolipid found in Gram-negative enteric bacteria. In Escherichia coli two gene products, RffH and RmlA, catalyze the same enzymatic reaction and are homologous in sequence; however, they are part of different operons and function in different pathways. We report the crystal structure of RffH bound to deoxythymidine triphosphate (dTTP), the phosphate donor, and Mg(2+), refined at 2.6 A to an R-factor of 22.3% (R(free) = 28.4%). The crystal structure of RffH shows a tetrameric enzyme best described as a dimer of dimers. Each monomer has an overall alpha/beta fold and consists of two domains, a larger nucleotide binding domain (residues 1-115, 222-291) and a smaller sugar-binding domain (116-221), with the active site located at the domain interface. The Mg(2+) ion is coordinated by two conserved aspartates and the alpha-phosphate of deoxythymidine triphosphate. Its location corresponds well to that in a structurally similar domain of N-acetylglucosamine-1-phosphate uridylyltransferase (GlmU). Analysis of the RffH, RmlA, and GlmU complexes with substrates and products provides an explanation for their different affinities for Mg(2+) and leads to a proposal for the dynamics along the reaction pathway.
==About this Structure==
==About this Structure==
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1MC3 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 MG and TTP as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Glucose-1-phosphate_thymidylyltransferase Glucose-1-phosphate thymidylyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.7.24 2.7.7.24] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1MC3 OCA].
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1MC3 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=MG:'>MG</scene> and <scene name='pdbligand=TTP:'>TTP</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Glucose-1-phosphate_thymidylyltransferase Glucose-1-phosphate thymidylyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.7.24 2.7.7.24] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1MC3 OCA].
==Reference==
==Reference==
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[[Category: Glucose-1-phosphate thymidylyltransferase]]
[[Category: Glucose-1-phosphate thymidylyltransferase]]
[[Category: Single protein]]
[[Category: Single protein]]
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[[Category: BSGI, Montreal-Kingston.Bacterial.Structural.Genomics.Initiative.]]
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[[Category: BSGI, Montreal-Kingston Bacterial Structural Genomics Initiative.]]
[[Category: Cygler, M.]]
[[Category: Cygler, M.]]
[[Category: Matte, A.]]
[[Category: Matte, A.]]
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[[Category: structural genomics]]
[[Category: structural genomics]]
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''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Nov 20 21:19:31 2007''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 13:53:51 2008''

Revision as of 11:53, 21 February 2008


1mc3, resolution 2.6Å

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CRYSTAL STRUCTURE OF RFFH

Overview

The enzyme glucose-1-phosphate thymidylyltransferase (RffH), the product of the rffh gene, catalyzes one of the steps in the synthesis of enterobacterial common antigen (ECA), a cell surface glycolipid found in Gram-negative enteric bacteria. In Escherichia coli two gene products, RffH and RmlA, catalyze the same enzymatic reaction and are homologous in sequence; however, they are part of different operons and function in different pathways. We report the crystal structure of RffH bound to deoxythymidine triphosphate (dTTP), the phosphate donor, and Mg(2+), refined at 2.6 A to an R-factor of 22.3% (R(free) = 28.4%). The crystal structure of RffH shows a tetrameric enzyme best described as a dimer of dimers. Each monomer has an overall alpha/beta fold and consists of two domains, a larger nucleotide binding domain (residues 1-115, 222-291) and a smaller sugar-binding domain (116-221), with the active site located at the domain interface. The Mg(2+) ion is coordinated by two conserved aspartates and the alpha-phosphate of deoxythymidine triphosphate. Its location corresponds well to that in a structurally similar domain of N-acetylglucosamine-1-phosphate uridylyltransferase (GlmU). Analysis of the RffH, RmlA, and GlmU complexes with substrates and products provides an explanation for their different affinities for Mg(2+) and leads to a proposal for the dynamics along the reaction pathway.

About this Structure

1MC3 is a Single protein structure of sequence from Escherichia coli with and as ligands. Active as Glucose-1-phosphate thymidylyltransferase, with EC number 2.7.7.24 Full crystallographic information is available from OCA.

Reference

Crystal structure of Escherichia coli glucose-1-phosphate thymidylyltransferase (RffH) complexed with dTTP and Mg2+., Sivaraman J, Sauve V, Matte A, Cygler M, J Biol Chem. 2002 Nov 15;277(46):44214-9. Epub 2002 Aug 8. PMID:12171937

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