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Journal of Bacteriology, December 2001, p. 7053-7057, Vol. 183, No. 24
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.24.7053-7057.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

The PPP-Family Protein Phosphatases PrpA and PrpB of Salmonella enterica Serovar Typhimurium Possess Distinct Biochemical Properties

Liang Shi,* David G. Kehres, and Michael E. Maguire

Department of Pharmacology, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106

Received 10 August 2001/Accepted 21 September 2001

Salmonella enterica serovar Typhimurium requires Mn2+, but only a few Mn2+-dependent enzymes have been identified from bacteria. To characterize Mn2+-dependent enzymes from serovar Typhimurium, two putative PPP-family protein phosphatase genes were cloned from serovar Typhimurium and named prpA and prpB. Their DNA-derived amino acid sequences showed 61% identity to the corresponding Escherichia coli proteins and 41% identity to each other. Each phosphatase was expressed in E. coli and purified to near electrophoretic homogeneity. Both PrpA and PrpB absolutely required a divalent metal for activity. As with other phosphatases of this class, Mn2+ had the highest affinity and stimulated the greatest activity. The apparent Ka of PrpA for Mn2+ of 65 µM was comparable to that for other bacterial phosphatases, but PrpB had a much higher affinity for Mn2+ (1.3 µM). The pH optima were pH 6.5 for PrpA and pH 8 for PrpB, while the optimal temperatures were 45 to 55°C for PrpA and 30 to 37°C for PrpB. Each phosphatase could hydrolyze phosphorylated serine, threonine, or tyrosine residues, but their relative specific activities varied with the specific substrate tested. These differences suggest that each phosphatase is used by serovar Typhimurium under different growth or environmental conditions such as temperature or acidity.

* Corresponding author. Mailing address: Department of Pharmacology, School of Medicine, Case Western Reserve University, 10900 Euclid Ave., Cleveland, OH 44106-4965. Phone: (216) 368-6187. Fax: (216) 368-3395. E-mail: lxs76{at}po.cwru.edu.

Journal of Bacteriology, December 2001, p. 7053-7057, Vol. 183, No. 24
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.24.7053-7057.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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