1m79: Difference between revisions

Revision as of 14:52, 21 February 2008

Candida albicans Dihydrofolate Reductase Complexed with Dihydro-Nicotinamide-Adenine-Dinucleotide Phosphate (NADPH) and 5-(4-methoxyphenoxy)-2,4-quinazolinediamine (GW1466)

OverviewOverview

The recent rise in systemic fungal infections has created a need for the development of new antifungal agents. As part of an effort to provide therapeutically effective inhibitors of fungal dihydrofolate reductase (DHFR), we have cloned, expressed, purified, crystallized, and determined the three-dimensional structure of Candida albicans DHFR. The 192-residue enzyme, which was expressed in Escherichia coli and purified by methotrexate affinity and cation exchange chromatography, was 27% identical to human DHFR. Crystals of C. albicans DHFR were grown as the holoenzyme complex and as a ternary complex containing a pyrroloquinazoline inhibitor. Both complexes crystallized with two molecules in the asymmetric unit in space group P21. The final structures had R-factors of 0.199 at 1.85-A resolution and 0.155 at 1.60-A resolution, respectively. The enzyme fold was similar to that of bacterial and vertebrate DHFR, and the binding of a nonselective diaminopyrroloquinazoline inhibitor and the interactions of NADPH with protein were typical of ligand binding to other DHFRs. However, the width of the active site cleft of C. albicans DHFR was significantly larger than that of the human enzyme, providing a basis for the design of potentially selective inhibitors.

About this StructureAbout this Structure

1M79 is a Single protein structure of sequence from Candida albicans with , and as ligands. Active as Dihydrofolate reductase, with EC number 1.5.1.3 Full crystallographic information is available from OCA.

ReferenceReference

X-ray crystallographic studies of Candida albicans dihydrofolate reductase. High resolution structures of the holoenzyme and an inhibited ternary complex., Whitlow M, Howard AJ, Stewart D, Hardman KD, Kuyper LF, Baccanari DP, Fling ME, Tansik RL, J Biol Chem. 1997 Nov 28;272(48):30289-98. PMID:9374515

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-[[Image:1m79.gif|left|200px]]<br /><applet load="1m79" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:1m79.gif|left|200px]]<br /><applet load="1m79" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="1m79, resolution 1.70&Aring;" />
 '''Candida albicans Dihydrofolate Reductase Complexed with Dihydro-Nicotinamide-Adenine-Dinucleotide Phosphate (NADPH) and 5-(4-methoxyphenoxy)-2,4-quinazolinediamine (GW1466)'''<br />
 ==Overview==
-The recent rise in systemic fungal infections has created a need for the, development of new antifungal agents. As part of an effort to provide, therapeutically effective inhibitors of fungal dihydrofolate reductase, (DHFR), we have cloned, expressed, purified, crystallized, and determined, the three-dimensional structure of Candida albicans DHFR. The 192-residue, enzyme, which was expressed in Escherichia coli and purified by, methotrexate affinity and cation exchange chromatography, was 27%, identical to human DHFR. Crystals of C. albicans DHFR were grown as the, holoenzyme complex and as a ternary complex containing a, pyrroloquinazoline inhibitor. Both complexes crystallized with two, molecules in the asymmetric unit in space group P21. The final structures, had R-factors of 0.199 at 1.85-A resolution and 0.155 at 1.60-A, resolution, respectively. The enzyme fold was similar to that of bacterial, and vertebrate DHFR, and the binding of a nonselective, diaminopyrroloquinazoline inhibitor and the interactions of NADPH with, protein were typical of ligand binding to other DHFRs. However, the width, of the active site cleft of C. albicans DHFR was significantly larger than, that of the human enzyme, providing a basis for the design of potentially, selective inhibitors.
+The recent rise in systemic fungal infections has created a need for the development of new antifungal agents. As part of an effort to provide therapeutically effective inhibitors of fungal dihydrofolate reductase (DHFR), we have cloned, expressed, purified, crystallized, and determined the three-dimensional structure of Candida albicans DHFR. The 192-residue enzyme, which was expressed in Escherichia coli and purified by methotrexate affinity and cation exchange chromatography, was 27% identical to human DHFR. Crystals of C. albicans DHFR were grown as the holoenzyme complex and as a ternary complex containing a pyrroloquinazoline inhibitor. Both complexes crystallized with two molecules in the asymmetric unit in space group P21. The final structures had R-factors of 0.199 at 1.85-A resolution and 0.155 at 1.60-A resolution, respectively. The enzyme fold was similar to that of bacterial and vertebrate DHFR, and the binding of a nonselective diaminopyrroloquinazoline inhibitor and the interactions of NADPH with protein were typical of ligand binding to other DHFRs. However, the width of the active site cleft of C. albicans DHFR was significantly larger than that of the human enzyme, providing a basis for the design of potentially selective inhibitors.
 ==About this Structure==
-M79 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Candida_albicans Candida albicans] with NDP, MQ1 and MES as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Dihydrofolate_reductase Dihydrofolate reductase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.5.1.3 1.5.1.3] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1M79 OCA].
+M79 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Candida_albicans Candida albicans] with <scene name='pdbligand=NDP:'>NDP</scene>, <scene name='pdbligand=MQ1:'>MQ1</scene> and <scene name='pdbligand=MES:'>MES</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Dihydrofolate_reductase Dihydrofolate reductase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.5.1.3 1.5.1.3] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1M79 OCA].
 ==Reference==
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 [[Category: Dihydrofolate reductase]]
 [[Category: Single protein]]
-[[Category: Howard, A.J.]]
+[[Category: Howard, A J.]]
-[[Category: Kuyper, L.F.]]
+[[Category: Kuyper, L F.]]
 [[Category: Whitlow, M.]]
 [[Category: MES]]
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 [[Category: reductase]]
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