Cloning, expression, purification and preliminary X-ray crystallographic studies of 2-methylisocitrate lyase from Salmonella typhimurium

Dhirendra K. Simanshu, P. S. Satheshkumar, S. Parthasarathy, H. S. Savithri, M. R N Murthy

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

In Salmonella typhimurium, propionate is oxidized to pyruvate via the 2-methylcitric acid cycle. The last step of this cycle, the cleavage of 2-methylisocitrate to succinate and pyruvate, is catalysed by 2-methylisocitrate lyase (EC 4.1.3.30). Methylisocitrate lyase (molecular weight 32 kDa) with a C-terminal polyhistidine affinity tag has been cloned and overexpressed in Escherichia coli and purified and crystallized under different conditions using the hanging-drop vapour-diffusion technique. Crystals belong to the orthogonal space group P212121, with unit-cell parameters a = 63.600, b = 100.670, c = 204.745 Å. A complete data set to 2.5 Å resolution has been collected using an image-plate detector system mounted on a rotating-anode X-ray generator.

Original languageEnglish (US)
Pages (from-to)2159-2161
Number of pages3
JournalActa Crystallographica Section D: Biological Crystallography
Volume58
Issue number12
DOIs
StatePublished - Dec 1 2002
Externally publishedYes

Fingerprint

pyruvates
salmonella
Salmonella
Cloning
Salmonella typhimurium
Pyruvic Acid
purification
Purification
Organism Cloning
X-Rays
X rays
cycles
Propionates
Succinic Acid
Escherichia
Escherichia coli
affinity
cleavage
molecular weight
Anodes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biophysics
  • Clinical Biochemistry
  • Structural Biology
  • Condensed Matter Physics

Cite this

Cloning, expression, purification and preliminary X-ray crystallographic studies of 2-methylisocitrate lyase from Salmonella typhimurium. / Simanshu, Dhirendra K.; Satheshkumar, P. S.; Parthasarathy, S.; Savithri, H. S.; Murthy, M. R N.

In: Acta Crystallographica Section D: Biological Crystallography, Vol. 58, No. 12, 01.12.2002, p. 2159-2161.

Research output: Contribution to journalArticle

Simanshu, Dhirendra K. ; Satheshkumar, P. S. ; Parthasarathy, S. ; Savithri, H. S. ; Murthy, M. R N. / Cloning, expression, purification and preliminary X-ray crystallographic studies of 2-methylisocitrate lyase from Salmonella typhimurium. In: Acta Crystallographica Section D: Biological Crystallography. 2002 ; Vol. 58, No. 12. pp. 2159-2161.
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AU - Savithri, H. S.

AU - Murthy, M. R N

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