3′-Phosphoadenosine 5′-Phosphosulfate (PAPS) Synthases, Naturally Fragile Enzymes Specifically Stabilized by Nucleotide Binding

Boom, Johannes van den; Heider, Dominik; Martin, Stephen R.; Pastore, Annalisa; Mueller, Jonathan Wolf

doi:10.1074/jbc.m111.325498

Cited by 40 publications

(58 citation statements)

References 38 publications

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“…A sequence alignment of 101 vertebrate and invertebrate sequences (15) was extended to plant, fungal, and yeast sequences of ATP sulfurylase. The alignment was created using Clustal W (16).…”

Section: Methodsmentioning

confidence: 99%

PAPSS2 Deficiency Causes Androgen Excess via Impaired DHEA Sulfation—In Vitro and in Vivo Studies in a Family Harboring Two Novel PAPSS2 Mutations

Oostdijk

Idkowiak

Mueller

et al. 2015

The Journal of Clinical Endocrinology &Amp; Metabolism

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Context:PAPSS2 (PAPS synthase 2) provides the universal sulfate donor PAPS (3′-phospho-adenosine-5′-phosphosulfate) to all human sulfotransferases, including SULT2A1, responsible for sulfation of the crucial androgen precursor dehydroepiandrosterone (DHEA). Impaired DHEA sulfation is thought to increase the conversion of DHEA toward active androgens, a proposition supported by the previous report of a girl with inactivating PAPSS2 mutations who presented with low serum DHEA sulfate and androgen excess, clinically manifesting with premature pubarche and early-onset polycystic ovary syndrome.Patients and Methods:We investigated a family harboring two novel PAPSS2 mutations, including two compound heterozygous brothers presenting with disproportionate short stature, low serum DHEA sulfate, but normal serum androgens. Patients and parents underwent a DHEA challenge test comprising frequent blood sampling and urine collection before and after 100 mg DHEA orally, with subsequent analysis of DHEA sulfation and androgen metabolism by mass spectrometry. The functional impact of the mutations was investigated in silico and in vitro.Results:We identified a novel PAPSS2 frameshift mutation, c.1371del, p.W462Cfs*3, resulting in complete disruption, and a novel missense mutation, c.809G>A, p.G270D, causing partial disruption of DHEA sulfation. Both patients and their mother, who was heterozygous for p.W462Cfs*3, showed increased 5α-reductase activity at baseline and significantly increased production of active androgens after DHEA intake. The mother had a history of oligomenorrhea and chronic anovulation that required clomiphene for ovulation induction.Conclusions:We provide direct in vivo evidence for the significant functional impact of mutant PAPSS2 on DHEA sulfation and androgen activation. Heterozygosity for PAPSS2 mutations can be associated with a phenotype resembling polycystic ovary syndrome.

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Section: Methodsmentioning

confidence: 99%

PAPSS2 Deficiency Causes Androgen Excess via Impaired DHEA Sulfation—In Vitro and in Vivo Studies in a Family Harboring Two Novel PAPSS2 Mutations

Oostdijk

Idkowiak

Mueller

et al. 2015

The Journal of Clinical Endocrinology &Amp; Metabolism

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“…Within the cell, strong product inhibition of the sulfurylase enzyme by APS would not be an obstacle if the next enzyme in the sulfate activation sequence (APS kinase) had a high affinity for APS. All vertebrates, and at least all sequenced invertebrates, including insects and worms, have bifunctional PAPS synthases in the form of a C-terminal ATP sulfurylase domain and an N-terminal APS kinase domain connected by a short irregular linker [11]. Intermediate channeling has been suggested for this type of PAPS synthase, as a possible way to circumvent APS inhibition [31], but this has been clearly disproved biochemically [13] and structurally [29] for human PAPSS1.…”

Section: Bifunctional Paps Synthases Are Stabilized By Apsmentioning

confidence: 99%

“…Recently, we made the unexpected observation that the two human isoforms of PAPS synthase differ remarkably in their stability towards chemically and thermally induced unfolding [11]. In a search for stabilizing substances for the PAPS synthases, we tested the substrates, intermediates and products of PAPS formation itself for their ability to stabilize these fragile proteins.…”

Section: Bifunctional Paps Synthases Are Stabilized By Apsmentioning

confidence: 99%

“…APS at 50 lM even caused changes in the overall form of the unfolding transitions, as it stabilized an unfolding intermediate by > 16°C in both PAPS synthases. From the analysis of several point mutants, the transition stabilization at higher APS concentrations was attributed to the unfolding of the sulfurylase domain of bifunctional PAPS synthases [11].…”

Section: Bifunctional Paps Synthases Are Stabilized By Apsmentioning

confidence: 99%

“…Generally, APS kinases are also inhibited by APS. Finally, APS has recently been recognized as a specific stabilizer of the human PAPS synthases, PAPS synthase 1 (PAPSS1) and PAPS synthase 2 (PAPSS2) [11], bifunctional enzymes that consist of ATP sulfurylase and APS kinase domains connected by a flexible linker. This review highlights the various regulatory roles of APS in the overall process of PAPS biosynthesis.…”

Section: Introductionmentioning

confidence: 99%

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Adenosine‐5′‐phosphosulfate – a multifaceted modulator of bifunctional 3′‐phospho‐adenosine‐5′‐phosphosulfate synthases and related enzymes

Mueller

Shafqat

2013

The FEBS Journal

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All sulfation reactions rely on active sulfate in the form of 3′-phospho-adenosine-5′-phosphosulfate (PAPS). In fungi, bacteria, and plants, the enzymes responsible for PAPS synthesis, ATP sulfurylase and adenosine-5′-phosphosulfate (APS) kinase, reside on separate polypeptide chains. In metazoans, however, bifunctional PAPS synthases catalyze the consecutive steps of sulfate activation by converting sulfate to PAPS via the intermediate APS. This intricate molecule and the related nucleotides PAPS and 3′-phospho-adenosine-5′-phosphate modulate the function of various enzymes from sulfation pathways, and these effects are summarized in this review. On the ATP sulfurylase domain that initially produces APS from sulfate and ATP, APS acts as a potent product inhibitor, being competitive with both ATP and sulfate. For the APS kinase domain that phosphorylates APS to PAPS, APS is an uncompetitive substrate inhibitor that can bind both at the ATP/ADP-binding site and the PAPS/APS-binding site. For human PAPS synthase 1, the steady-state concentration of APS has been modelled to be 1.6 μm, but this may increase up to 60 μm under conditions of sulfate excess. It is noteworthy that the APS concentration for maximal APS kinase activity is 15 μm. Finally, we recognized APS as a highly specific stabilizer of bifunctional PAPS synthases. APS most likely stabilizes the APS kinase part of these proteins by forming a dead-end enzyme–ADP–APS complex at APS concentrations between 0.5 and 5 μm; at higher concentrations, APS may bind to the catalytic centers of ATP sulfurylase. Based on the assumption that cellular concentrations of APS fluctuate within this range, APS can therefore be regarded as a key modulator of PAPS synthase functions.

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Small World: A Plant Perspective on Human Sulfate Activation

Mueller

O’Neill

Shafqat

2015

Proceedings of the International Plant Sulfur Workshop

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3′-Phosphoadenosine 5′-Phosphosulfate (PAPS) Synthases, Naturally Fragile Enzymes Specifically Stabilized by Nucleotide Binding

Cited by 40 publications

References 38 publications

PAPSS2 Deficiency Causes Androgen Excess via Impaired DHEA Sulfation—In Vitro and in Vivo Studies in a Family Harboring Two Novel PAPSS2 Mutations

PAPSS2 Deficiency Causes Androgen Excess via Impaired DHEA Sulfation—In Vitro and in Vivo Studies in a Family Harboring Two Novel PAPSS2 Mutations

Adenosine‐5′‐phosphosulfate – a multifaceted modulator of bifunctional 3′‐phospho‐adenosine‐5′‐phosphosulfate synthases and related enzymes

Small World: A Plant Perspective on Human Sulfate Activation

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