Characterization of recombinant Francisella tularensis acid phosphatase A

• Published in: Protein expression and purification 2006, Vol.45 (1), p.132-141
• Main Authors: Reilly, Thomas J., Felts, Richard L., Henzl, Michael T., Calcutt, Michael J., Tanner, John J.
• Format: Article
• Language: English
• Subjects: Acid Phosphatase - chemistry; Acid Phosphatase - isolation & purification; Acid Phosphatase - metabolism; Acid phosphatase A; Bacterial Proteins - chemistry; Bacterial Proteins - isolation & purification; Bacterial Proteins - metabolism; Burst-inhibiting acid phosphatase; Francisella tularensis; Francisella tularensis - enzymology; Hydrogen-Ion Concentration; Kinetics; Molecular Weight; Phosphomonoesterase; Recombinant Proteins - chemistry; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Substrate Specificity; TRAP
• ISSN: 1046-5928; 1096-0279
• DOI: 10.1016/j.pep.2005.05.001

Francisella tularensis is the etiologic agent of the potentially fatal human disease tularemia and is capable of survival and multiplication within professional phagocytes of the host. While the mechanisms that allow intracellular survival of the bacterium are only now beginning to be elucidated at the molecular level, previous work demonstrated that F. tularensis produces copious levels of an acid phosphatase which in crude and purified form affected the dose-dependent abrogation of the respiratory burst of stimulated neutrophils. The work presented here was undertaken to provide a source of recombinant F. tularensis acid phosphatase for detailed biochemical, biological, and structural studies. Results from this work are consistent with the ability to generate milligram amounts of recombinant enzyme whose attributes are demonstrably equivalent to those of the native enzyme. Such properties include molecular mass, broad substrate specificity, sensitivity and resistance to various inhibitors, pH optimum, and reactivity with rabbit polyclonal antibody to the native enzyme.