Oxaloacetate 4-carboxy-lyase from Pseudomanas ovalis Chester

Horton, Ashley A; Kornberg, H. L.

doi:10.1016/0926-6569(64)90236-6

Cited by 14 publications

(11 citation statements)

References 7 publications

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“…In addition, the enzyme appears to be constitutive. The enzyme is, therefore, not of the same type as that found in A. aerogenes (20) and appears to be more closely related to the OAA decarboxylases observed in other bacteria (2,9,10,12,13,16,17). However, it also differs from these in not being stimulated by Mg2+ or Mn2+ and being only slightly inhibited by EDTA.…”

Section: Resultsmentioning

confidence: 61%

Citrate Metabolism in Aerobacter cloacae

O'Brien

Geisler

1974

J Bacteriol

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Section: Resultsmentioning

confidence: 61%

Citrate Metabolism in Aerobacter cloacae

O'Brien

Geisler

1974

J Bacteriol

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“…For assay of the enzyme in crude extracts or when the effect of NAD was tested, the lactate dehydrogenase and NADH were omitted, NAD and OAA were added as specified, and the pyruvate formed was determined after termination of the reaction with 20 mM ethylenediaminetetraacetate, as described below (method II). In some experiments the reaction was followed spectrophotometrically by determining the rate of decrease in absorption at 262 nm concomitant with the decarboxylation of OAA (16) (method III). The activity was corrected for the nonenzymatic decarboxylation of OAA, which did not exceed 5% of total readings.…”

Section: Methodsmentioning

confidence: 99%

Purification and Regulatory Properties of the Oxaloacetate Decarboxylase of Acetobacter xylinum

et al. 1978

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The oxaloacetate (OAA) decarboxylase (EC 4.1.1.3) activity of Acetobacter xylinum cells grown on glucose or glycerol is the same as that of cells grown on intermediates of the citrate cycle. The enzyme was purified 92-fold from extracts, and its molecular weight was determined to be 100,000 by gel filtration. Initial velocity studies revealed marked positive cooperativity for OAA (Hill coefficient [nH] = 1.8; So.5 = 21 mM). The affinity of the enzyme for OAA was markedly increased upon addition of nicotinamide adenine dinucleotide (NAD), NAD phosphate (NADP), and some other pyridine nucleotides. SO.5(OAA) decreased to 1 mM but nH and Vm. were unchanged. Saturation kinetics for the pyridine nucleotides were hyperbolic, and a half-maximal effect was obtained with 8 ,uM NAD and 30 uM NADP. The enzyme also catalyzed the exchange of '4CO2 into OAA but not the net carboxylation of pyruvate. Exchange activity, too, exhibited sigmoidal kinetics for OAA and was strongly stimulated by NAD at low substrate concentrations. The enzyme was inhibited by acetate competitively with respect to OAA. The KI for acetate (12 mM) was well within the physiological range of this compound inside the cell. The regulatory properties of the decarboxylase with respect to OAA cooperativity, NAD activation, and acetate inhibition were retained in situ within permeabilized cells. These properties seem to provide for a control mechanism which could insure the maintenance of OAA and the citrate cycle during growth of cells on glucose and, conversely, the required supply of pyruvate during growth on intermediates of the citrate cycle.

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“…The β subunit catalyses the decarboxylation of biotin and pumps two sodium ions out of the cell membrane [4][5][6], whereas the γ subunit is mainly responsible for stabilization of the whole complex [7,8]. Several cytosolic OADs have been characterized in various bacterial species [10][11][12]. Several cytosolic OADs have been characterized in various bacterial species [10][11][12].…”

Section: Introductionmentioning

confidence: 99%

Crystal structures of Cg1458 reveal a catalytic lid domain and a common catalytic mechanism for the FAH family

Ran

Gao

Marsh

et al. 2012

Biochemical Journal

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Cg1458 was recently characterized as a novel soluble oxaloacetate decarboxylase. However, sequence alignment identified that Cg1458 has no similarity with other oxaloacetate decarboxylases and instead belongs to the FAH (fumarylacetoacetate hydrolase) family. Differences in the function of Cg1458 and other FAH proteins may suggest a different catalytic mechanism. To help elucidate the catalytic mechanism of Cg1458, crystal structures of Cg1458 in both the open and closed conformations have been determined for the first time up to a resolution of 1.9 Å (1 Å=0.1 nm) and 2.0 Å respectively. Comparison of both structures and detailed biochemical studies confirmed the presence of a catalytic lid domain which is missing in the native enzyme structure. In this lid domain, a glutamic acid-histidine dyad was found to be critical in mediating enzymatic catalysis. On the basis of structural modelling and comparison, as well as large-scale sequence alignment studies, we further determined that the catalytic mechanism of Cg1458 is actually through a glutamic acid-histidine-water triad, and this catalytic triad is common among FAH family proteins that catalyse the cleavage of the C-C bond of the substrate. Two sequence motifs, HxxE and Hxx…xxE have been identified as the basis for this mechanism.

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Oxaloacetate 4-carboxy-lyase from Pseudomanas ovalis Chester

Cited by 14 publications

References 7 publications

Citrate Metabolism in Aerobacter cloacae

Citrate Metabolism in Aerobacter cloacae

Purification and Regulatory Properties of the Oxaloacetate Decarboxylase of Acetobacter xylinum

Crystal structures of Cg1458 reveal a catalytic lid domain and a common catalytic mechanism for the FAH family

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