L-ribulose-5-phosphate 4-epimerase

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L-ribulose-phosphate 4-epimerase
Ribulose Epimerase Pymol.jpg
Identifiers
EC no. 5.1.3.4
CAS no. 9024-19-5
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MetaCyc metabolic pathway
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In enzymology, a L-ribulose-5-phosphate 4-epimerase (EC 5.1.3.4) is an enzyme that catalyzes the interconversion of ribulose 5-phosphate and xylulose 5-phosphate in the oxidative phase of the Pentose phosphate pathway. [1]

Contents

L-ribulose 5-phosphate D-xylulose 5-phosphate

This enzyme has a molecular mass of 102 kDa and is believed to be composed of four identical 25.5 kDa subunits. It belongs to the family of isomerases, specifically those racemases and epimerases acting on carbohydrates and derivatives. [2] The systematic name of this enzyme class is L-ribulose-5-phosphate 4-epimerase. Other names in common use include phosphoribulose isomerase, ribulose phosphate 4-epimerase, L-ribulose-phosphate 4-epimerase, L-ribulose 5-phosphate 4-epimerase, AraD, and L-Ru5P. This enzyme participates in pentose and glucuronate interconversions and ascorbate and aldarate metabolism.

Enzyme Mechanism

L-Ribulose 5-phosphate 4-epimerase catalyzes the epimerization of L-ribulose 5-phosphate to D-xylulose 5-phosphate by retro-aldol cleavage and subsequent aldol reaction. The proposed mechanism involves the abstraction of the proton from the hydroxyl group on C-4, followed by cleavage of the bond between C-3 and C-4 to give a metal-stabilized acetone enediolate and a glycolaldehyde phosphate fragment. The C–C bond of glycolaldehyde phosphate is then rotated 180°, and the C–C bond between C-3 and C-4 is regenerated to give inversion of stereochemistry at C-4. [3]

This mechanism is contested by a possible alternative dehydration reaction scheme. The literature [2] [3] favors the aldol mechanism for two reasons. First, the retro-aldol cleavage mechanism is analogous to the reaction catalyzed by L-fuculose-phosphate aldolase which has high levels of sequence similarity with L-ribulose-5-phosphate 4-epimerase. Second, the analysis of 13C and deuterium kinetic isotope effects points toward the aldol mechanism. It has been reported that there is little to no difference in the deuterium isotope effects at C-3 and C-4, suggesting that these C–H bonds are not broken during epimerization. [3] Changes in isotope effect at C-3 would be expected for the dehydration mechanism, because the breaking of the C–H bond is the rate-limiting step in this mechanism and substituting the C-3 hydrogen with deuterium would significantly alter the rate. At the same time there are significantly large 13C isotope effects, suggesting rate-limiting C–C bond breakage, as expected with the aldol mechanism. [3]

Structure

The structure is homo-tetrameric and displays C4 symmetry. [4] Each protein subunit has a single domain consisting of a central β sheet flanked on either side by layers of α-helix. A central β-sheet is formed from nine β-strands (b1-b9) and is predominantly antiparallel except between strands b7 and b8. The eight α-helices of the structure form two layers on either side of the central β-sheet. The active site is identified by the position of the catalytic zinc residue and is located at the interface between two adjacent subunits. Asp76, His95, His97, and His171act as the metal-binding residues. A remarkable feature of the structure is that it shows a very close resemblance to that of L-fuculose-phosphate aldolase. [2] This is consistent with the notion that both enzymes belong to a superfamily of epimerases/aldolases that catalyze carbon-carbon bond cleavage reactions via a metal-stabilized enolate intermediate.

Biological Function

Ribulose 5-phosphate 4-epimerase is found on the well studied L-arabinose operon. This operon consists of eight genes araA-araH with the gene for Ribulose 5-phosphate 4-epimerase called araD. The arabinose system enables the take up the pentose L-arabinose, and then the conversion of intracellular arabinose in three steps catalyzed by the products of the araB, araA, araD genes to D-xylulose-5-phosphate. [5]

GeneProtein
AraAIsomerase
AraBRibulokinase
AraCRegulatory
AraDEpimerase
AraEUptake
AraFUptake
AraGUptake
AraHUptake

Evolution

L-Ribulose-5-phosphate 4-epimerase and L-fuculose-1-phosphate (L-Fuc1P) aldolase are evolutionarily related enzymes that display 26% sequence identity and a very high degree of structural similarity. [2] They both employ a divalent cation in the stabilization of an enolate during catalysis, and both are able to deprotonate the C-4 hydroxyl group of a phosphoketose substrate. Despite these many similarities, subtle distinctions are present which allow the enzymes to catalyze two seemingly different reactions and to accommodate substrates differing greatly in the position of the phosphate (C-5 vs C-1). [6]

Related Research Articles

Isomerases are a general class of enzymes that convert a molecule from one isomer to another. Isomerases facilitate intramolecular rearrangements in which bonds are broken and formed. The general form of such a reaction is as follows:

A tetrose is a monosaccharide with 4 carbon atoms. They have either an aldehyde functional group in position 1 (aldotetroses) or a ketone functional group in position 2 (ketotetroses).

<span class="mw-page-title-main">Ribulose</span> Monosaccharide with five carbon atoms and a ketone functional group

Ribulose is a ketopentose — a monosaccharide containing five carbon atoms, and including a ketone functional group. It has chemical formula C5H10O5. Two enantiomers are possible, d-ribulose and l-ribulose. d-Ribulose is the diastereomer of d-xylulose.

<span class="mw-page-title-main">Transketolase</span> Enzyme involved in metabolic pathways

Transketolase is an enzyme that, in humans, is encoded by the TKT gene. It participates in both the pentose phosphate pathway in all organisms and the Calvin cycle of photosynthesis. Transketolase catalyzes two important reactions, which operate in opposite directions in these two pathways. In the first reaction of the non-oxidative pentose phosphate pathway, the cofactor thiamine diphosphate accepts a 2-carbon fragment from a 5-carbon ketose (D-xylulose-5-P), then transfers this fragment to a 5-carbon aldose (D-ribose-5-P) to form a 7-carbon ketose (sedoheptulose-7-P). The abstraction of two carbons from D-xylulose-5-P yields the 3-carbon aldose glyceraldehyde-3-P. In the Calvin cycle, transketolase catalyzes the reverse reaction, the conversion of sedoheptulose-7-P and glyceraldehyde-3-P to pentoses, the aldose D-ribose-5-P and the ketose D-xylulose-5-P.

The L-arabinose operon, also called the ara or araBAD operon, is an operon required for the breakdown of the five-carbon sugar L-arabinose in Escherichia coli. The L-arabinose operon contains three structural genes: araB, araA, araD, which encode for three metabolic enzymes that are required for the metabolism of L-arabinose. AraB (ribulokinase), AraA, AraD produced by these genes catalyse conversion of L-arabinose to an intermediate of the pentose phosphate pathway, D-xylulose-5-phosphate.

<span class="mw-page-title-main">Serine hydroxymethyltransferase</span>

Serine hydroxymethyltransferase (SHMT) is a pyridoxal phosphate (PLP) (Vitamin B6) dependent enzyme (EC 2.1.2.1) which plays an important role in cellular one-carbon pathways by catalyzing the reversible, simultaneous conversions of L-serine to glycine and tetrahydrofolate (THF) to 5,10-Methylenetetrahydrofolate (5,10-CH2-THF). This reaction provides the largest part of the one-carbon units available to the cell.

<span class="mw-page-title-main">6-Phosphogluconate dehydrogenase</span> Class of enzymes

6-Phosphogluconate dehydrogenase (6PGD) is an enzyme in the pentose phosphate pathway. It forms ribulose 5-phosphate from 6-phosphogluconate:

<span class="mw-page-title-main">Transaldolase</span> Enzyme family

Transaldolase is an enzyme of the non-oxidative phase of the pentose phosphate pathway. In humans, transaldolase is encoded by the TALDO1 gene.

<span class="mw-page-title-main">Xylulose 5-phosphate</span> Chemical compound

D-Xylulose 5-phosphate (D-xylulose-5-P) is an intermediate in the pentose phosphate pathway. It is a ketose sugar formed from ribulose-5-phosphate by ribulose-5-phosphate epimerase. In the non-oxidative branch of the pentose phosphate pathway, xylulose-5-phosphate acts as a donor of two-carbon ketone groups in transketolase reactions.

<span class="mw-page-title-main">Phosphopentose epimerase</span>

Phosphopentose epimerase encoded by the RPE gene is a metalloprotein that catalyzes the interconversion between D-ribulose 5-phosphate and D-xylulose 5-phosphate.

<span class="mw-page-title-main">Xylose metabolism</span>

D-Xylose is a five-carbon aldose that can be catabolized or metabolized into useful products by a variety of organisms.

<span class="mw-page-title-main">Phosphogluconate dehydrogenase (decarboxylating)</span>

In enzymology, a phosphogluconate dehydrogenase (decarboxylating) (EC 1.1.1.44) is an enzyme that catalyzes the chemical reaction

<span class="mw-page-title-main">L-arabinose isomerase</span>

In enzymology, a L-arabinose isomerase is an enzyme that catalyzes the chemical reaction

In enzymology, a L-ribulose-5-phosphate 3-epimerase is an enzyme that catalyzes the chemical reaction

<span class="mw-page-title-main">Ribose-5-phosphate isomerase</span>

Ribose-5-phosphate isomerase (Rpi) encoded by the RPIA gene is an enzyme that catalyzes the conversion between ribose-5-phosphate (R5P) and ribulose-5-phosphate (Ru5P). It is a member of a larger class of isomerases which catalyze the interconversion of chemical isomers. It plays a vital role in biochemical metabolism in both the pentose phosphate pathway and the Calvin cycle. The systematic name of this enzyme class is D-ribose-5-phosphate aldose-ketose-isomerase.

In enzymology, an UDP-arabinose 4-epimerase is an enzyme that catalyzes the chemical reaction

<span class="mw-page-title-main">2-Dehydro-3-deoxy-phosphogluconate aldolase</span> Class of enzymes

The enzyme 2-dehydro-3-deoxy-phosphogluconate aldolase, commonly known as KDPG aldolase, catalyzes the chemical reaction

The enzyme L-fuculose-phosphate aldolase (EC 4.1.2.17) catalyzes the chemical reaction

In enzymology, a D-ribulokinase is an enzyme that catalyzes the chemical reaction

<span class="mw-page-title-main">Ribulokinase</span>

In enzymology, a ribulokinase is an enzyme that catalyzes the chemical reaction

References

  1. Englesberg, E.; R.L. Anderson; R. Weinberg (29 January 1962). "L-Arabinose-Sensitive, L-Ribulose 5-phosphate 4-Epimerase-Deficient Mutants of Escherichia coli". Journal of Bacteriology. 84 (137): 137–46. doi:10.1128/JB.84.1.137-146.1962. PMC   277779 . PMID   13890280.
  2. 1 2 3 4 Yu, Luo; Jomy Samuel; Steven C Mosimann; Jeffrey E Lee; Martin E Tanner; Natalie CJ Strynadka (2001). "The Structure of Ribulose-5-Phosphate 4-Epimerase: An Aldolase-like Platform for Epimerization". Biochemistry. 40 (49): 14763–14771. CiteSeerX   10.1.1.510.5360 . doi:10.1021/bi0112513. PMID   11732895.
  3. 1 2 3 4 Lee LV, Vu MV, Cleland WW (April 2000). "13C and deuterium isotope effects suggest an aldol cleavage mechanism for L-ribulose-5-phosphate 4-epimerase". Biochemistry. 39 (16): 4808–20. CiteSeerX   10.1.1.537.5159 . doi:10.1021/bi992894+. PMID   10769138.
  4. Andersson, Arnold; Gunter Schneider (15 May 1995). "Purification and preliminary X-ray crystallographic studies of recombinant ~-ribulose-5-phosphate 4-epimerase from Escherichia coli". Protein Science. 4 (12): 1648–1650. doi:10.1002/pro.5560040823. PMC   2143197 . PMID   8520491.
  5. Schlelf, Robert (December 2000). "Regulation of the L-arabinose operon of Escherichia coli". Trends in Genetics. 16 (12): 559–564. doi:10.1016/S0168-9525(00)02153-3. PMID   11102706.[ dead link ]
  6. Samuel, Jomy; Yu Luo; Paul M Morgan; Natalie CJ Strynadka; Martin E Tanner (9 November 2001). "Catalysis and binding in L-Ribulose 5-Phosphate 4-Epimerase: A Comparison with L-Fuculose Phosphate Aldolase". Biochemistry. 40 (49): 14772–14780. doi:10.1021/bi011252v. PMID   11732896.

Further reading