Biochemical characterization of aspartyl phosphate phosphatase interaction with a phosphorylated response regulator and its inhibition by a pentapeptide.

The Journal of Biological Chemistry
Shu IshikawaMarta Perego

Abstract

The RapA and RapB proteins are aspartyl phosphate phosphatases that specifically dephosphorylate the Spo0F approximately P intermediate response regulator of the phosphorelay signal transduction system for sporulation initiation in Bacillus subtilis. The approximately 48-kDa His-tag derivative proteins were purified by metal affinity chromatography, and their molecular and biochemical characteristics were studied. RapA and RapB were found to be dimers in solution. Enzymatic activity was strongly dependent upon maintaining reducing conditions during purification and storage. RapA phosphatase activity on Spo0F approximately P is inhibited in vivo by a pentapeptide generated from the phrA gene. Native gel assays demonstrated that the RapA dimer forms a stable complex with two molecules of Spo0F approximately P or with its PhrA pentapeptide inhibitor. The pentapeptide was shown to displace Spo0F approximately P from a preformed complex with RapA. The structural organization of Rap phosphatases in tetratricopeptide repeats provides insights on the mechanisms of RapA interaction with its substrate and its inhibitor.

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Citations

Aug 5, 2003·Journal of Bacteriology·Sophie StephensonMarta Perego
Feb 4, 2003·Journal of Bacteriology·Lee Kroos, Janine R Maddock
Sep 18, 2004·Journal of Bacteriology·Xiaoqiang WuJames W Golden
Jun 22, 2005·Journal of Bacteriology·Cristina BongiorniMarta Perego
Dec 31, 2005·Journal of Bacteriology·Cristina BongiorniMarta Perego
Feb 25, 2011·PLoS Biology·Vijay ParasharMatthew B Neiditch
Jan 5, 2012·PLoS Biology·Melinda D Baker, Matthew B Neiditch
Feb 6, 2010·Current Opinion in Microbiology·Ruth E Silversmith
Nov 24, 2004·Current Opinion in Microbiology·Patrick J Piggot, David W Hilbert
Dec 25, 2009·FEMS Microbiology Reviews·Daniel López, Roberto Kolter
Aug 3, 2016·PLoS Pathogens·Stéphane PerchatSylvie Nessler
Jan 19, 2017·Nature·Zohar ErezRotem Sorek
Oct 3, 2017·Environmental Microbiology·Fernanda FazionDidier Lereclus
Feb 20, 2020·Journal of Bacteriology·Abel Verdugo-FuentesMayra de la Torre
Jul 1, 2020·Journal of Bacteriology·Aisha T Burton, Daniel B Kearns
Dec 26, 2002·The Journal of Biological Chemistry·Vanessa RioliEmer S Ferro
Sep 10, 2017·Annual Review of Microbiology·James A Hoch
Mar 7, 2020·Nature Communications·Heiko BabelIlka B Bischofs

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