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Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of succinyl-diaminopimelate desuccinylase (Rv1202, DapE) from Mycobacterium tuberculosis

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  • Linda Reinhard (maiden name Schuldt)
  • ,
  • Jochen Mueller-Dieckmann
  • ,
  • Manfred S Weiss
  • Molekylærbiologisk Institut
Succinyl-diaminopimelate desuccinylase from Mycobacterium tuberculosis (DapE, Rv1202) has been cloned, heterologously expressed in Escherichia coli and purified using standard chromatographic techniques. Diffraction-quality crystals were obtained at acidic pH from ammonium sulfate and PEG and diffraction data were collected from two crystals to resolutions of 2.40 and 2.58 Å, respectively. The crystals belonged to the monoclinic space group P2(1), with unit-cell parameters a = 79.7, b = 76.0, c = 82.9 Å, β = 119°. The most probable content of the asymmetric unit was two molecules of DapE, which would correspond to a solvent content of 56%. Both examined crystals turned out to be pseudo-merohedrally twinned, with twin operator -h, -k, h + l and twin fractions of approximately 0.46 and 0.16, respectively.
OriginalsprogEngelsk
TidsskriftActa Crystallographica. Section F: Structural Biology and Crystallization Communications Online
Vol/bind68
NummerPt 9
Sider (fra-til)1089-93
Antal sider5
ISSN2053-230X
DOI
StatusUdgivet - 2012

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