Purification and some properties of liver adenylylsulfate kinase

Hommes, F. A.; Moss, Linda; Touchton, J. T.

doi:10.1016/0304-4165(87)90022-5

Cited by 16 publications

(4 citation statements)

References 15 publications

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“…enzyme activities and does not need to form larger oligomeric complexes for activity. However, it is still possible that under certain conditions the enzyme forms aggregates, as previously described (Geller et al, 1987;Matsuo et al, 1987;Hommes et al, 1987), which may or may not represent the physiologically relevant state.…”

Section: Resultsmentioning

confidence: 99%

“…However, large molecular weight aggregates containing enzyme activity were also eluted under certain conditions. Hommes et al (1987) have reported that rat liver APS kinase is a 58-kDa protein comprised of four equal subunits. In earlier studies, rat chondrosarcoma ATP sulfurylase and APS kinase copurified over 2000-fold through S300 gel filtration, hydroxylapatite, and ATP-affinity chromatography (Geller et al, 1987), suggesting that the activities are inseparable.…”

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confidence: 99%

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Rat Chondrosarcoma ATP Sulfurylase and Adenosine 5'-Phosphosulfate Kinase Reside on a Single Bifunctional Protein

Lyle¹,

Stanczak²,

Ng³

et al. 1994

Biochemistry

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The sulfate-activation pathway consists of the sequential action of ATP sulfurylase (ATP: sulfate adenylyltransferase, EC 2.7.7.4) and adenosine 5'-phosphosulfate kinase ATP:adenylylsulfate 3'-phosphotransferase, EC 2.7.1.25). Both sulfurylase and kinase from rat chondrosarcoma were copurified through substrate affinity chromatography using stable analogs of APS (adenosine 5'-phosphosulfate) and PAPS (3'-phosphoadenosine 5'-phosphosulfate). A 56-kDa protein, containing both activities, was then purified to apparent homogeneity through reversed-phase chromatography and observed on denaturing polyacrylamide gels. The molecular mass of the native active unit containing both activities in the purified preparation corresponded to approximately 60 kDa by analytical gel filtration. Coincident binding and elution of ATP sulfurylase and APS kinase by immunoaffinity chromatography, using polyclonal serum generated against the 56-kDa protein, also demonstrated that the enzyme contains both activities. Lastly, a single N-terminal amino acid sequence was obtained from the 56-kDa band isolated by gel electrophoresis. These results all suggest that ATP sulfurylase and APS kinase from rat chondrosarcoma residue on a single bifunctional protein.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Rat Chondrosarcoma ATP Sulfurylase and Adenosine 5'-Phosphosulfate Kinase Reside on a Single Bifunctional Protein

Lyle¹,

Stanczak²,

Ng³

et al. 1994

Biochemistry

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“…Interestingly, the P. chrysogenum APS kinase (13,14) as well as the Arabidopsis thaliana (15), human (9), and rat (16,17) enzymes have been reported to exhibit pronounced uncompetitive substrate inhibition by APS. Kinetic data suggest that the inhibitory complex results from formation of the E⅐ADP⅐APS complex (Scheme 1).…”

Section: Atp-mg ϩ Somentioning

confidence: 99%

Structural Mechanism for Substrate Inhibition of the Adenosine 5′-Phosphosulfate Kinase Domain of Human 3′-Phosphoadenosine 5′-Phosphosulfate Synthetase 1 and Its Ramifications for Enzyme Regulation

Sekulić

Konrad

Lavie

2007

Journal of Biological Chemistry

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In mammals, the universal sulfuryl group donor molecule 3-phosphoadenosine 5-phosphosulfate (PAPS) is synthesized in two steps by a bifunctional enzyme called PAPS synthetase. The APS kinase domain of PAPS synthetase catalyzes the second step in which APS, the product of the ATP-sulfurylase domain, is phosphorylated on its 3-hydroxyl group to yield PAPS. The substrate APS acts as a strong uncompetitive inhibitor of the APS kinase reaction. We generated truncated and point mutants of the APS kinase domain that are active but devoid of substrate inhibition. Structural analysis of these mutant enzymes reveals the intrasubunit rearrangements that occur upon substrate binding. We also observe intersubunit rearrangements in this dimeric enzyme that result in asymmetry between the two monomers. Our work elucidates the structural elements required for the ability of the substrate APS to inhibit the reaction at micromolar concentrations. Because the ATPsulfurylase domain of PAPS synthetase influences these elements in the APS kinase domain, we propose that this could be a communication mechanism between the two domains of the bifunctional enzyme.Enzyme inhibitors are molecules that bind to enzymes and decrease their activity. Usually, we think of inhibitors as molecules that mimic the substrates, products, or transition state or fit in an allosteric site and, once bound, arrest the enzyme in its catalytic cycle or slow it down. However, even the true substrates (or products) of the reaction can act as inhibitors. In such cases, enzyme kinetic measurements show pronounced deviations from the hyperbolic dependence of velocity on substrate concentration. Substrate inhibition is usually observed only at high substrate concentrations, whereas at low substrate concentrations the kinetics follow simple Michaelis-Menten behavior.Substrate or product inhibition phenomena are widely found in nature and believed to be important for metabolic feedback regulation processes (1-3). However, the underlying mechanisms are complex and structurally poorly characterized. In this work, we shed light on one such system present in adenosine 5Ј-phosphosulfate (APS) 4 kinase. APS kinase is an evolutionarily conserved enzyme catalyzing the second step in the formation of 3Ј-phosphoadenosine 5Ј-phosphosulfate (PAPS), the universal sulfuryl donor in biological systems. In the first reaction of PAPS formation, inorganic sulfate is converted to APS by ATP-sulfurylase. In the second reaction, APS kinase phosphorylates APS at the 3Ј-hydroxyl of the ribose to produce PAPS. ATP-MgPAPS is used as a substrate for sulfotransferases, which sulfonate a wide rage of molecules, including proteins, glycosaminoglycans, steroid hormones, and xenobiotics (4). ATP-sulfurylase and APS kinase are separate enzymes in bacteria, yeast, fungi, and plants, but are found on a single polypeptide chain in the animal kingdom (5). In its bifunctional form, called PAPS synthetase or PAPSS, each domain retains the same fold as in the homologous monofunctional enzyme (6 -8). The s...

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“…In Penicillium chrysogenum (Lansdon et al, 2002;MacRae et al, 2000;Renosto et al, 1985), Saccharomyces cerevisiae (Schriek & Schwenn, 1986;, Arabidopsis thaliana (Lillig et al, 2001) and Escherichia coli (Satishchandran et al, 1992;Satishchandran & Markham, 1989;Schriek & Schwenn, 1986), APS kinase is found as a single-domain peptide that displays solely APS kinase activity. However, APS kinase can also be found as part of a bifunctional enzyme that also contains ATP sulfurylase activity as in Aquifex aeolicus (Hanna et al, 2002;Yu et al, 2007), rats (Hommes et al, 1987;Lyle et al, 1994;Yu et al, 1989) and humans (Harjes et al, 2005;Lansdon et al, 2004). Regardless of its source, all structural data gathered to date reveal the active form of APS kinase to be a homodimer.…”

Section: Introductionmentioning

confidence: 99%

Structure of the two-domain hexameric APS kinase fromThiobacillus denitrificans: structural basis for the absence of ATP sulfurylase activity

Segel

Fisher

2009

Acta Crystallogr D Biol Cryst

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The Tbd_0210 gene of the chemolithotrophic bacterium Thiobacillus denitrificans is annotated to encode a 60.5 kDa bifunctional enzyme with ATP sulfurylase and APS kinase activity. This putative bifunctional enzyme was cloned, expressed and structurally characterized. The 2.95 A resolution X-ray crystal structure reported here revealed a hexameric assembly with D(3) symmetry. Each subunit contains a large N-terminal sulfurylase-like domain and a C-terminal APS kinase domain reminiscent of the two-domain fungal ATP sulfurylases of Penicillium chrysogenum and Saccharomyces cerevisiae, which also exhibit a hexameric assembly. However, the T. denitrificans enzyme exhibits numerous structural and sequence differences in the N-terminal domain that render it inactive with respect to ATP sulfurylase activity. Surprisingly, the C-terminal domain does indeed display APS kinase activity, indicating that this gene product is a true APS kinase. Therefore, these results provide the first structural insights into a unique hexameric APS kinase that contains a nonfunctional ATP sulfurylase-like domain of unknown function.

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Purification and some properties of liver adenylylsulfate kinase

Cited by 16 publications

References 15 publications

Rat Chondrosarcoma ATP Sulfurylase and Adenosine 5'-Phosphosulfate Kinase Reside on a Single Bifunctional Protein

Rat Chondrosarcoma ATP Sulfurylase and Adenosine 5'-Phosphosulfate Kinase Reside on a Single Bifunctional Protein

Structural Mechanism for Substrate Inhibition of the Adenosine 5′-Phosphosulfate Kinase Domain of Human 3′-Phosphoadenosine 5′-Phosphosulfate Synthetase 1 and Its Ramifications for Enzyme Regulation

Structure of the two-domain hexameric APS kinase fromThiobacillus denitrificans: structural basis for the absence of ATP sulfurylase activity

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