Expression, purification and preliminary crystallographic analysis of Mycobacterium tuberculosis CysQ, a phosphatase involved in sulfur metabolism

Anna I. Erickson, Reta D. Sarsam, Andrew J Fisher

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1 Citation (Scopus)

Abstract

CysQ is part of the sulfur-activation pathway that dephosphorylates 3′-phosphoadenosine 5′-monophosphate (PAP) to regenerate adenosine 5′-monophosphate (AMP) and free phosphate. PAP is the product of sulfate-transfer reactions from sulfotransferases that use the universal sulfate donor 3′-phosphoadenosine 5′-phosphosulfate (PAPS). In some organisms PAP is also the product of PAPS reductases that reduce sulfate from PAPS to sulfite. CysQ from Mycobacterium tuberculosis, which plays an important role in the biosynthesis of sulfated glycoconjugates, was successfully purified and crystallized in 24% PEG 1500, 20% glycerol. X-ray diffraction data were collected to 1.7 Å resolution using a synchrotron-radiation source. Crystals grew in the orthorhombic space group P212121, with unit-cell parameters a = 40.3, b = 57.9, c = 101.7 Å and with one monomer per asymmetric unit.

Original languageEnglish (US)
Pages (from-to)750-753
Number of pages4
JournalActa Crystallographica Section F:Structural Biology Communications
Volume70
Issue number6
DOIs
StatePublished - 2014

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Phosphoadenosine Phosphosulfate
tuberculosis
phosphatases
metabolism
Mycobacterium tuberculosis
Phosphoric Monoester Hydrolases
Sulfur
Metabolism
purification
Sulfates
Purification
sulfates
sulfur
Sulfotransferases
sulfites
adenosines
Sulfites
Synchrotrons
biosynthesis
Glycoconjugates

Keywords

  • 3′,5′-bisphosphate nucleotidase
  • 3′-phosphoadenosine 5′-phosphate
  • CysQ
  • monophosphatase
  • Mycobacterium tuberculosis
  • PAP
  • Rv2131c

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Genetics
  • Structural Biology
  • Condensed Matter Physics
  • Medicine(all)

Cite this

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title = "Expression, purification and preliminary crystallographic analysis of Mycobacterium tuberculosis CysQ, a phosphatase involved in sulfur metabolism",
abstract = "CysQ is part of the sulfur-activation pathway that dephosphorylates 3′-phosphoadenosine 5′-monophosphate (PAP) to regenerate adenosine 5′-monophosphate (AMP) and free phosphate. PAP is the product of sulfate-transfer reactions from sulfotransferases that use the universal sulfate donor 3′-phosphoadenosine 5′-phosphosulfate (PAPS). In some organisms PAP is also the product of PAPS reductases that reduce sulfate from PAPS to sulfite. CysQ from Mycobacterium tuberculosis, which plays an important role in the biosynthesis of sulfated glycoconjugates, was successfully purified and crystallized in 24{\%} PEG 1500, 20{\%} glycerol. X-ray diffraction data were collected to 1.7 {\AA} resolution using a synchrotron-radiation source. Crystals grew in the orthorhombic space group P212121, with unit-cell parameters a = 40.3, b = 57.9, c = 101.7 {\AA} and with one monomer per asymmetric unit.",
keywords = "3′,5′-bisphosphate nucleotidase, 3′-phosphoadenosine 5′-phosphate, CysQ, monophosphatase, Mycobacterium tuberculosis, PAP, Rv2131c",
author = "Erickson, {Anna I.} and Sarsam, {Reta D.} and Fisher, {Andrew J}",
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T1 - Expression, purification and preliminary crystallographic analysis of Mycobacterium tuberculosis CysQ, a phosphatase involved in sulfur metabolism

AU - Erickson, Anna I.

AU - Sarsam, Reta D.

AU - Fisher, Andrew J

PY - 2014

Y1 - 2014

N2 - CysQ is part of the sulfur-activation pathway that dephosphorylates 3′-phosphoadenosine 5′-monophosphate (PAP) to regenerate adenosine 5′-monophosphate (AMP) and free phosphate. PAP is the product of sulfate-transfer reactions from sulfotransferases that use the universal sulfate donor 3′-phosphoadenosine 5′-phosphosulfate (PAPS). In some organisms PAP is also the product of PAPS reductases that reduce sulfate from PAPS to sulfite. CysQ from Mycobacterium tuberculosis, which plays an important role in the biosynthesis of sulfated glycoconjugates, was successfully purified and crystallized in 24% PEG 1500, 20% glycerol. X-ray diffraction data were collected to 1.7 Å resolution using a synchrotron-radiation source. Crystals grew in the orthorhombic space group P212121, with unit-cell parameters a = 40.3, b = 57.9, c = 101.7 Å and with one monomer per asymmetric unit.

AB - CysQ is part of the sulfur-activation pathway that dephosphorylates 3′-phosphoadenosine 5′-monophosphate (PAP) to regenerate adenosine 5′-monophosphate (AMP) and free phosphate. PAP is the product of sulfate-transfer reactions from sulfotransferases that use the universal sulfate donor 3′-phosphoadenosine 5′-phosphosulfate (PAPS). In some organisms PAP is also the product of PAPS reductases that reduce sulfate from PAPS to sulfite. CysQ from Mycobacterium tuberculosis, which plays an important role in the biosynthesis of sulfated glycoconjugates, was successfully purified and crystallized in 24% PEG 1500, 20% glycerol. X-ray diffraction data were collected to 1.7 Å resolution using a synchrotron-radiation source. Crystals grew in the orthorhombic space group P212121, with unit-cell parameters a = 40.3, b = 57.9, c = 101.7 Å and with one monomer per asymmetric unit.

KW - 3′,5′-bisphosphate nucleotidase

KW - 3′-phosphoadenosine 5′-phosphate

KW - CysQ

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KW - Mycobacterium tuberculosis

KW - PAP

KW - Rv2131c

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