Crystallization and preliminary X-ray diffraction analysis of shikimate kinase from Mycobacterium tuberculosis in complex with MgADP

Acta Crystallogr D Biol Crystallogr. 2001 Dec;57(Pt 12):1870-1. doi: 10.1107/s0907444901014032. Epub 2001 Nov 21.

Abstract

Shikimate kinase (SK) from Mycobacterium tuberculosis (Mt) was overexpressed in Escherichia coli, purified and cocrystallized with MgADP in hanging drops using the vapor-diffusion procedure with PEG 4000 and 2-propanol as precipitants at pH 7.5. The crystal of MtSK-MgADP, which diffracted to 2.2 A resolution, belonged to space group P3(2)21 or P3(1)21, with unit-cell parameters a = b = 64.01, c = 92.41 A. There was one MtSK molecule in the asymmetric unit. Molecular-replacement trials with the crystal structure of SK from Erwinia chrysanthemi (PDB code 1shk) and adenylate kinase (PDB code 1ake) as search models were not successful. Heavy-atom derivative screening is in progress.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Diphosphate / chemistry*
  • Cloning, Molecular
  • Crystallization
  • Crystallography, X-Ray
  • Data Collection
  • Models, Molecular
  • Mycobacterium tuberculosis / enzymology*
  • Phosphotransferases (Alcohol Group Acceptor) / chemistry*
  • Phosphotransferases (Alcohol Group Acceptor) / genetics
  • Protein Conformation
  • Recombinant Proteins / chemistry

Substances

  • Recombinant Proteins
  • Adenosine Diphosphate
  • Phosphotransferases (Alcohol Group Acceptor)
  • shikimate kinase

Associated data

  • PDB/1AKE
  • PDB/1SHK