1qmg: Difference between revisions

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ACETOHYDROXYACID ISOMEROREDUCTASE COMPLEXED WITH ITS REACTION PRODUCT DIHYDROXY-METHYLVALERATE, MANGANESE AND ADP-RIBOSE.

OverviewOverview

Acetohydroxyacid isomeroreductase catalyses a two-step reaction composed of an alkyl migration followed by an NADPH-dependent reduction. Both steps require a divalent cation and the first step has a strong preference for magnesium. Manganese ions are highly unfavourable to the reaction: only 3% residual activity is observed in the presence of this cation. Acetohydroxyacid isomeroreductase has been crystallized with its substrate, 2-aceto-2-hydroxybutyrate (AHB), Mn(2+) and NADPH. The 1.6 A resolution electron-density map showed the reaction product (2,3-dihydroxy-3-methylvalerate, DHMV) and a density corresponding to (phospho)-ADP-ribose instead of the whole NADP(+). This is one of the few structures of an enzyme complexed with its reaction product. The structure of this complex was refined to an R factor of 19.3% and an R(free) of 22.5%. The overall structure of the enzyme is very similar to that of the complex with the reaction-intermediate analogue IpOHA [N-hydroxy-N-isopropyloxamate; Biou et al. (1997), EMBO J. 16, 3405-3415]. However, the active site shows some differences: the nicotinamide is cleaved and the surrounding amino acids have rearranged accordingly. Comparison between the structures corresponding to the reaction intermediate and to the end of the reaction allowed the proposal of a reaction scheme. Taking this result into account, the enzyme was crystallized with Ni(2+) and Zn(2+), for which only 0.02% residual activity were measured; however, the crystals of AHB/Zn/NADPH and of AHB/Ni/NADPH also contain the reaction product. Moreover, mass-spectrometry measurements confirmed the -cleavage of nicotinamide.

About this StructureAbout this Structure

1QMG is a Single protein structure of sequence from Spinacia oleracea with , , and as ligands. Active as Ketol-acid reductoisomerase, with EC number 1.1.1.86 Full crystallographic information is available from OCA.

ReferenceReference

Structure of spinach acetohydroxyacid isomeroreductase complexed with its reaction product dihydroxymethylvalerate, manganese and (phospho)-ADP-ribose., Thomazeau K, Dumas R, Halgand F, Forest E, Douce R, Biou V, Acta Crystallogr D Biol Crystallogr. 2000 Apr;56(Pt 4):389-97. PMID:10739911

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-[[Image:1qmg.jpg|left|200px]]<br /><applet load="1qmg" size="450" color="white" frame="true" align="right" spinBox="true"
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 '''ACETOHYDROXYACID ISOMEROREDUCTASE COMPLEXED WITH ITS REACTION PRODUCT DIHYDROXY-METHYLVALERATE, MANGANESE AND ADP-RIBOSE.'''<br />
 ==Overview==
-Acetohydroxyacid isomeroreductase catalyses a two-step reaction composed, of an alkyl migration followed by an NADPH-dependent reduction. Both steps, require a divalent cation and the first step has a strong preference for, magnesium. Manganese ions are highly unfavourable to the reaction: only 3%, residual activity is observed in the presence of this cation., Acetohydroxyacid isomeroreductase has been crystallized with its, substrate, 2-aceto-2-hydroxybutyrate (AHB), Mn(2+) and NADPH. The 1.6 A, resolution electron-density map showed the reaction product, (2,3-dihydroxy-3-methylvalerate, DHMV) and a density corresponding to, (phospho)-ADP-ribose instead of the whole NADP(+). This is one of the few, structures of an enzyme complexed with its reaction product. The structure, of this complex was refined to an R factor of 19.3% and an R(free) of, 22.5%. The overall structure of the enzyme is very similar to that of the, complex with the reaction-intermediate analogue IpOHA, [N-hydroxy-N-isopropyloxamate; Biou et al. (1997), EMBO J. 16, 3405-3415]., However, the active site shows some differences: the nicotinamide is, cleaved and the surrounding amino acids have rearranged accordingly., Comparison between the structures corresponding to the reaction, intermediate and to the end of the reaction allowed the proposal of a, reaction scheme. Taking this result into account, the enzyme was, crystallized with Ni(2+) and Zn(2+), for which only 0.02% residual, activity were measured; however, the crystals of AHB/Zn/NADPH and of, AHB/Ni/NADPH also contain the reaction product. Moreover, mass-spectrometry measurements confirmed the -cleavage of nicotinamide.
+Acetohydroxyacid isomeroreductase catalyses a two-step reaction composed of an alkyl migration followed by an NADPH-dependent reduction. Both steps require a divalent cation and the first step has a strong preference for magnesium. Manganese ions are highly unfavourable to the reaction: only 3% residual activity is observed in the presence of this cation. Acetohydroxyacid isomeroreductase has been crystallized with its substrate, 2-aceto-2-hydroxybutyrate (AHB), Mn(2+) and NADPH. The 1.6 A resolution electron-density map showed the reaction product (2,3-dihydroxy-3-methylvalerate, DHMV) and a density corresponding to (phospho)-ADP-ribose instead of the whole NADP(+). This is one of the few structures of an enzyme complexed with its reaction product. The structure of this complex was refined to an R factor of 19.3% and an R(free) of 22.5%. The overall structure of the enzyme is very similar to that of the complex with the reaction-intermediate analogue IpOHA [N-hydroxy-N-isopropyloxamate; Biou et al. (1997), EMBO J. 16, 3405-3415]. However, the active site shows some differences: the nicotinamide is cleaved and the surrounding amino acids have rearranged accordingly. Comparison between the structures corresponding to the reaction intermediate and to the end of the reaction allowed the proposal of a reaction scheme. Taking this result into account, the enzyme was crystallized with Ni(2+) and Zn(2+), for which only 0.02% residual activity were measured; however, the crystals of AHB/Zn/NADPH and of AHB/Ni/NADPH also contain the reaction product. Moreover, mass-spectrometry measurements confirmed the -cleavage of nicotinamide.
 ==About this Structure==
-QMG is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Spinacia_oleracea Spinacia oleracea] with MN, SO4, DMV and APX as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Ketol-acid_reductoisomerase Ketol-acid reductoisomerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.1.86 1.1.1.86] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1QMG OCA].
+QMG is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Spinacia_oleracea Spinacia oleracea] with <scene name='pdbligand=MN:'>MN</scene>, <scene name='pdbligand=SO4:'>SO4</scene>, <scene name='pdbligand=DMV:'>DMV</scene> and <scene name='pdbligand=APX:'>APX</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Ketol-acid_reductoisomerase Ketol-acid reductoisomerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.1.86 1.1.1.86] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1QMG OCA].
 ==Reference==
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 [[Category: reaction product]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 00:48:34 2007''
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