1887

Abstract

Among the few regulatory proteins encoded by is HPr kinase/phosphatase (HPrK/P), the key regulator of carbon metabolism in low-GC Gram-positive bacteria. The corresponding gene, , and the gene encoding the target protein HPr, , were overexpressed. analysis of the purified proteins confirmed ATP-dependent phosphorylation of HPr by HPrK/P. In contrast to HPrK/P of , which is by default a phosphatase and needs high ATP concentrations for kinase activity, the enzyme exhibits kinase activity at very low ATP concentrations and depends on P for phosphatase activity. This inverted control of enzymic activity may result from the adaptation to very different ecological niches. While the standard activities of HPrK/P from and other Gram-positive bacteria differ, they are both modulated by the concentration of ATP, P and glycolytic intermediates. Site-directed mutagenesis of a potential ATP-binding site and of the HPrK/P signature sequence resulted in four different activity classes: (i) inactive proteins, (ii) enzymes with reduced kinase and phosphatase activities, (iii) enzymes that had lost phosphatase, but not kinase activity, and (iv) enzymes that exhibited increased phosphatase activity.

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2002-10-01
2020-11-28
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