Expression, purification and characterization of UvrD2 helicase from Mycobacterium tuberculosis

Protein Expr Purif. 2010 Feb;69(2):215-8. doi: 10.1016/j.pep.2009.09.006. Epub 2009 Sep 18.

Abstract

Helicases appear to be important for genome stability of dormant Mycobacterium tuberculosis responsible for latent tuberculosis infection and big proportion of active disease cases caused by reactivation. It was demonstrated that in both M. tuberculosis and Mycobacterium smegmatis, a helicase termed UvrD2 is essential for bacterial growth making it a promising target to fight tuberculosis. In many cases expression of soluble and active mycobacterial proteins in Escherichia coli is a complicated issue. In this work we for the first time report a non-trivial expression procedure in E. coli, leading to soluble UvrD2 from M. tuberculosis which possesses DNA-dependent ATP-ase activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / metabolism
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Cloning, Molecular / methods
  • DNA / metabolism
  • DNA Helicases / genetics
  • DNA Helicases / metabolism*
  • Humans
  • Latent Tuberculosis / enzymology
  • Latent Tuberculosis / microbiology
  • Mycobacterium tuberculosis / enzymology*

Substances

  • Bacterial Proteins
  • DNA
  • Adenosine Triphosphatases
  • DNA Helicases