HisB from Mycobacterium tuberculosis: Cloning, overexpression in Mycobacterium smegmatis, purification, crystallization and preliminary X-ray crystallographic analysis

Mohammad Syed Ahangar, Yogesh Khandokar, Nazia Nasir, Rajan Vyas, Bichitra K. Biswal

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7 Citations (Scopus)

Abstract

HisB, encoded by open reading frame Rv1601, possesses enzymatic activity as an imidazoleglycerol-phosphate dehydratase in the histidine-biosynthetic pathway of Mycobacterium tuberculosis. A recombinant form of HisB was crystallized in three crystal forms: crystals grown using 20% PEG 1500 as a precipitant belonged to either the cubic space group P432 or the tetragonal space group P4, while an orthorhombic crystal form belonging to space group P21212 was obtained using 15% PEG 5000 and 10 mM MnCl2 as precipitant. The structure of HisB in the orthorhombic crystal form was solved by the molecular-replacement method using the crystal structure of its Arabidopsis thaliana counterpart, which shares 47% sequence identity with Rv1601, as the search model.

Original languageEnglish
Pages (from-to)1451-1456
Number of pages6
JournalActa Crystallographica Section F: Structural Biology Communications
Volume67
Issue number11
DOIs
Publication statusPublished - Nov 2011
Externally publishedYes

Keywords

  • HisB
  • Mycobacterium tuberculosis
  • Rv1601

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