Steady-State Kinetics of Cabbage Histidinol Dehydrogenase

Kheirolomoom, Azadeh; Mano, Junichi; Nagai, Atsuko; Ogawa, Atsuko; Iwasaki, Genji; Ohta, Daisaku

doi:10.1006/abbi.1994.1337

Cited by 22 publications

(19 citation statements)

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“…Spectra taken in the presence of added UDP-glucose and NAD ϩ indicate that the purified enzyme does not contain stoichiometric amounts of bound NAD ϩ or NADH. The kinetic mechanism of the bovine liver UDP-glucose dehydrogenase and the similar enzyme histidinol dehydrogenase (which also catalyzes the 2-fold oxidation of an alcohol to an acid without release of an aldehyde intermediate) have been investigated previously (42)(43)(44)(45). In both cases the results indicated that a bi-uni-uni-bi ping-pong mechanism was followed in which the alcohol was bound first and the acid was released last (Fig.…”

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confidence: 69%

“…Dialysis of the inactivated enzyme against inhibitor-free buffer for 48 h did not result in the restoration of any measurable activity, indicating that the process is irreversible. An electrospray mass spectrum of the inactivated enzyme showed that the mass of the protein increased from 45 decreased the rate of inactivation, suggesting that the inhibitor is active site-directed (see the following section).…”

Section: Fig 3 Uv-visible Spectra Of Udpgdhmentioning

confidence: 99%

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Properties and Kinetic Analysis of UDP-glucose Dehydrogenase from Group A Streptococci

Campbell

Sala

Rijn

et al. 1997

Journal of Biological Chemistry

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UDP-glucuronic acid is used by many pathogenic bacteria in the construction of an antiphagocytic capsule that is required for virulence. The enzyme UDP-glucose dehydrogenase catalyzes the NAD ؉ -dependent 2-fold oxidation of UDP-glucose and provides a source of the acid. In the present study the recombinant dehydrogenase from group A streptococci has been purified and found to be active as a monomer. The enzyme contains no chromophoric cofactors, and its activity is unaffected by the presence of EDTA or carbonyl-trapping reagents. Initial velocity and product inhibition kinetic patterns are consistent with a bi-uni-uni-bi ping-pong mechanism in which UDP-glucose is bound first and UDPglucuronate is released last. UDP-xylose was found to be a competitive inhibitor (K i , 2.7 M) of the enzyme. The enzyme is irreversibly inactivated by uridine 5 -diphosphate-chloroacetol due to the alkylation of an active site cysteine thiol. The apparent second order rate constant for the inhibition (k i /K i ) was found to be 2 ؋ 10 3 mM ؊1 min ؊1 . Incubation with the truncated compound, chloroacetol phosphate, resulted in no detectable inactivation when tested under comparable conditions. This supports the notion that uridine 5 -diphosphate-chloroacetol is bound in the place of UDP-glucose and is not simply acting as a nonspecific alkylating agent.The enzyme UDP-glucose dehydrogenase (UDPGDH) 1 catalyzes the NAD ϩ -dependent oxidation of UDP-glucose to UDPglucuronic acid (Fig. 1). It belongs to a small group of dehydrogenases that are able to carry out the 2-fold oxidation of an alcohol to an acid without the release of an aldehyde intermediate (1). Much of the work to date has focused on the properties and mechanism of the beef liver enzyme, and relatively little is known about the enzyme purified from bacterial sources (2-4). In many strains of bacteria that act as human pathogens, UDPGDH provides the UDP-glucuronic acid required for the construction of an antiphagocytic capsular polysaccharide. It is well established that the formation of the capsule is required for virulence (5, 6), and it is thought that the capsule enables the bacteria to evade the host's immune system (7,8). Group A and C streptococci are mammalian pathogens that use UDPGDH in the synthesis of a capsule composed of hyaluronic acid (a polysaccharide consisting of alternating glucuronic acid and N-acetylglucosamine residues) (9, 10). Many of the known strains of Streptococcus pneumoniae also use UDP-glucuronic acid in the construction of their polysaccharide capsule (11), and it has recently been shown that UDPGDH is required for capsule production in S. pneumoniae type 3 (12). The encapsulated Escherichia coli K5 is also known to use the enzyme for a similar purpose (2, 13). The hasB gene that encodes for UDPGDH in group A streptococci (Streptococcus pyogenes) has been cloned and overexpressed in Escherichia coli (14). Its gene product shares 57% sequence identity and 74% sequence similarity with the S. pneumoniae enzyme (15, 16). These properties make th...

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confidence: 69%

Section: Fig 3 Uv-visible Spectra Of Udpgdhmentioning

confidence: 99%

Properties and Kinetic Analysis of UDP-glucose Dehydrogenase from Group A Streptococci

Campbell

Sala

Rijn

et al. 1997

Journal of Biological Chemistry

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“…Overall, the enzyme catalyzes a four-electron oxidation reaction. A catalytic mechanism has been proposed for the conversion of L-histidinol to L-histidine, which involves two consecutive oxidation reactions accompanied by a reduction of two NAD ϩ molecules according to a Bi-Uni-Uni-Bi kinetic mechanism (17)(18)(19). On the basis of the available data, a catalytic mechanism has been proposed (14) (Fig.…”

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confidence: 99%

Mechanism of action and NAD ⁺ -binding mode revealed by the crystal structure of l -histidinol dehydrogenase

Barbosa

Sivaraman

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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The histidine biosynthetic pathway is an ancient one found in bacteria, archaebacteria, fungi, and plants that converts 5-phosphoribosyl 1-pyrophosphate to L-histidine in 10 enzymatic reactions. This pathway provided a paradigm for the operon, transcriptional regulation of gene expression, and feedback inhibition of a pathway. L-histidinol dehydrogenase (HisD, EC 1.1.1.23) catalyzes the last two steps in the biosynthesis of L-histidine: sequential NAD-dependent oxidations of L-histidinol to L-histidinaldehyde and then to L-histidine. HisD functions as a homodimer and requires the presence of one Zn 2؉ cation per monomer. We have determined the three-dimensional structure of Escherichia coli HisD in the apo state as well as complexes with substrate, Zn 2؉ , and NAD ؉ (best resolution is 1.7 Å). Each monomer is made of four domains, whereas the intertwined dimer possibly results from domain swapping. Two domains display a very similar incomplete Rossmann fold that suggests an ancient event of gene duplication. Residues from both monomers form the active site. Zn 2؉ plays a crucial role in substrate binding but is not directly involved in catalysis. The active site residue His-327 participates in acid-base catalysis, whereas Glu-326 activates a water molecule. NAD ؉ binds weakly to one of the Rossmann fold domains in a manner different from that previously observed for other proteins having a Rossmann fold.

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“…This compound, in turn, undergoes seven additional enzymatic reactions leading to histidine, the last two of which are catalyzed by histidinol-dehydrogenase (HisD) (Figure 1) i.e. the double oxidation of histidinol to histidine, through the intermediate histidinal, concomitant to the reduction of two NAD+ molecules, with a Bi-Uni-Uni-Bi kinetic mechanism [26,27]. …”

Section: Resultsmentioning

confidence: 99%

The role of gene fusions in the evolution of metabolic pathways: the histidine biosynthesis case

Fani¹,

Brilli²,

Fondi³

et al. 2007

BMC Evol Biol

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Background: Histidine biosynthesis is one of the best characterized anabolic pathways. There is a large body of genetic and biochemical information available, including operon structure, gene expression, and increasingly larger sequence databases. For over forty years this pathway has been the subject of extensive studies, mainly in Escherichia coli and Salmonella enterica, in both of which details of histidine biosynthesis appear to be identical. In these two enterobacteria the pathway is unbranched, includes a number of unusual reactions, and consists of nine intermediates; his genes are arranged in a compact operon (hisGDC [NB]HAF [IE]), with three of them (hisNB, hisD and hisIE) coding for bifunctional enzymes. We performed a detailed analysis of his gene fusions in available genomes to understand the role of gene fusions in shaping this pathway.

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Steady-State Kinetics of Cabbage Histidinol Dehydrogenase

Cited by 22 publications

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Properties and Kinetic Analysis of UDP-glucose Dehydrogenase from Group A Streptococci

Properties and Kinetic Analysis of UDP-glucose Dehydrogenase from Group A Streptococci

Mechanism of action and NAD ⁺ -binding mode revealed by the crystal structure of l -histidinol dehydrogenase

The role of gene fusions in the evolution of metabolic pathways: the histidine biosynthesis case

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Steady-State Kinetics of Cabbage Histidinol Dehydrogenase

Cited by 22 publications

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Properties and Kinetic Analysis of UDP-glucose Dehydrogenase from Group A Streptococci

Properties and Kinetic Analysis of UDP-glucose Dehydrogenase from Group A Streptococci

Mechanism of action and NAD + -binding mode revealed by the crystal structure of l -histidinol dehydrogenase

The role of gene fusions in the evolution of metabolic pathways: the histidine biosynthesis case

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Mechanism of action and NAD ⁺ -binding mode revealed by the crystal structure of l -histidinol dehydrogenase