Crystal structure of ATP sulfurylase from Penicillium chrysogenum

Insights into the allosteric regulation of sulfate assimilation

I. J. MacRae, I. H. Segel, Andrew J Fisher

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

ATP sulfurylase from Penicillium chrysogenum is an allosterically regulated enzyme composed of six identical 63.7 kDa subunits (573 residues). The C-terminal allosteric domain of each subunit is homologous to APS kinase. In the presence of APS, the enzyme crystallized in the orthorhombic space group (I222) with unit cell parameters of a = 135.7 Å, b = 162.1 Å, and c = 273.0 Å. The X-ray structure at 2.8 Å resolution established that the hexameric enzyme is a dimer of triads in the shape of an oblate ellipsoid 140 Å diameter × 70 Å. Each subunit is divided into a discreet N-terminal domain, a central catalytic domain, and a C-terminal allosteric domain. Two molecules of APS bound per subunit clearly identify the catalytic and allosteric domains. The sequence 197QXRN200 is largely responsible for anchoring the phosphosulfate group of APS at the active site of the catalytic domain. The specificity of the catalytic site for adenine nucleotides is established by specific hydrogen bonds to the protein main chain. APS was bound to the allosteric site through sequence-specific interactions with amino acid side chains that are conserved in true APS kinase. Within a given triad, the allosteric domain of one subunit interacts with the catalytic domain of another. There are also allosteric - allosteric, allosteric - N-terminal, and catalytic - catalytic domain interactions across the triad interface. The overall interactions - each subunit with four others - provide stability to the hexamer as well as a way to propagate a concerted allosteric transition. The structure presented here is believed to be the R state. A solvent channel, 15-70 Å wide exists along the 3-fold axis, but substrates have access to the catalytic site only from the external medium. On the other hand, a surface "trench" links each catalytic site in one triad with an allosteric site in the other triad. This trench may be a vestigial feature of a bifunctional ("PAPS synthetase") ancestor of fungal ATP sulfurylase.

Original languageEnglish (US)
Pages (from-to)6795-6804
Number of pages10
JournalBiochemistry
Volume40
Issue number23
DOIs
StatePublished - Jun 12 2001

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Sulfate Adenylyltransferase
Allosteric Regulation
Penicillium chrysogenum
adenylylsulfate kinase
Sulfates
Catalytic Domain
Crystal structure
Enzymes
Adenine Nucleotides
Dimers
Allosteric Site
Hydrogen bonds
Amino Acids
X rays
Molecules
Substrates
Proteins
Hydrogen

ASJC Scopus subject areas

  • Biochemistry

Cite this

Crystal structure of ATP sulfurylase from Penicillium chrysogenum : Insights into the allosteric regulation of sulfate assimilation. / MacRae, I. J.; Segel, I. H.; Fisher, Andrew J.

In: Biochemistry, Vol. 40, No. 23, 12.06.2001, p. 6795-6804.

Research output: Contribution to journalArticle

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title = "Crystal structure of ATP sulfurylase from Penicillium chrysogenum: Insights into the allosteric regulation of sulfate assimilation",
abstract = "ATP sulfurylase from Penicillium chrysogenum is an allosterically regulated enzyme composed of six identical 63.7 kDa subunits (573 residues). The C-terminal allosteric domain of each subunit is homologous to APS kinase. In the presence of APS, the enzyme crystallized in the orthorhombic space group (I222) with unit cell parameters of a = 135.7 {\AA}, b = 162.1 {\AA}, and c = 273.0 {\AA}. The X-ray structure at 2.8 {\AA} resolution established that the hexameric enzyme is a dimer of triads in the shape of an oblate ellipsoid 140 {\AA} diameter × 70 {\AA}. Each subunit is divided into a discreet N-terminal domain, a central catalytic domain, and a C-terminal allosteric domain. Two molecules of APS bound per subunit clearly identify the catalytic and allosteric domains. The sequence 197QXRN200 is largely responsible for anchoring the phosphosulfate group of APS at the active site of the catalytic domain. The specificity of the catalytic site for adenine nucleotides is established by specific hydrogen bonds to the protein main chain. APS was bound to the allosteric site through sequence-specific interactions with amino acid side chains that are conserved in true APS kinase. Within a given triad, the allosteric domain of one subunit interacts with the catalytic domain of another. There are also allosteric - allosteric, allosteric - N-terminal, and catalytic - catalytic domain interactions across the triad interface. The overall interactions - each subunit with four others - provide stability to the hexamer as well as a way to propagate a concerted allosteric transition. The structure presented here is believed to be the R state. A solvent channel, 15-70 {\AA} wide exists along the 3-fold axis, but substrates have access to the catalytic site only from the external medium. On the other hand, a surface {"}trench{"} links each catalytic site in one triad with an allosteric site in the other triad. This trench may be a vestigial feature of a bifunctional ({"}PAPS synthetase{"}) ancestor of fungal ATP sulfurylase.",
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N2 - ATP sulfurylase from Penicillium chrysogenum is an allosterically regulated enzyme composed of six identical 63.7 kDa subunits (573 residues). The C-terminal allosteric domain of each subunit is homologous to APS kinase. In the presence of APS, the enzyme crystallized in the orthorhombic space group (I222) with unit cell parameters of a = 135.7 Å, b = 162.1 Å, and c = 273.0 Å. The X-ray structure at 2.8 Å resolution established that the hexameric enzyme is a dimer of triads in the shape of an oblate ellipsoid 140 Å diameter × 70 Å. Each subunit is divided into a discreet N-terminal domain, a central catalytic domain, and a C-terminal allosteric domain. Two molecules of APS bound per subunit clearly identify the catalytic and allosteric domains. The sequence 197QXRN200 is largely responsible for anchoring the phosphosulfate group of APS at the active site of the catalytic domain. The specificity of the catalytic site for adenine nucleotides is established by specific hydrogen bonds to the protein main chain. APS was bound to the allosteric site through sequence-specific interactions with amino acid side chains that are conserved in true APS kinase. Within a given triad, the allosteric domain of one subunit interacts with the catalytic domain of another. There are also allosteric - allosteric, allosteric - N-terminal, and catalytic - catalytic domain interactions across the triad interface. The overall interactions - each subunit with four others - provide stability to the hexamer as well as a way to propagate a concerted allosteric transition. The structure presented here is believed to be the R state. A solvent channel, 15-70 Å wide exists along the 3-fold axis, but substrates have access to the catalytic site only from the external medium. On the other hand, a surface "trench" links each catalytic site in one triad with an allosteric site in the other triad. This trench may be a vestigial feature of a bifunctional ("PAPS synthetase") ancestor of fungal ATP sulfurylase.

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