Biochemical characterization of the class B acid phosphatase (AphA) of Escherichia coli MG1655

Claudio Passariello, Costantino Forleo, Vanna Micheli, Serena Schippa, Rosalida Leone, Stefano Mangani, Maria Cristina Thaller, Gian Maria Rossolini

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The AphA enzyme of Escherichia coli, a molecular class B periplasmic phosphatase that belongs to the DDDD superfamily of phosphohydrolases, was purified and subjected to biochemical characterization. Kinetic analysis with several substrates revealed that the enzyme essentially behaves as a broad-spectrum nucleotidase highly active on 3′- and 5′- mononucleotides and monodeoxynucleotides, but not active on cyclic nucleotides, or nucleotides di- and triphosphate. Mononucleotides are degraded to nucleosides, and AphA apparently does not exhibit any nucleotide phosphomutase activity. However, it can transphosphorylate nucleosides in the presence of phosphate donors. Kinetic properties of AphA are consistent with structural data, and suggest a role for the hydrophobic pocket present in the active site crevice, made by residues Phe 56, Leu71, Trp77 and Tyr193, in conferring preferential substrate specificity by accommodating compounds with aromatic rings. AphA was inhibited by several chelating agents, including EDTA, EGTA, 1,10-phenanthroline and dipicolinic acid, with EDTA being apparently the most powerful inhibitor.

Original languageEnglish
Pages (from-to)13-19
Number of pages7
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1764
Issue number1
DOIs
Publication statusPublished - Jan 2006

Fingerprint

Acid Phosphatase
Escherichia coli
nucleotidase
Nucleosides
Phosphoric Monoester Hydrolases
Phosphotransferases (Phosphomutases)
Edetic Acid
Nucleotides
Kinetics
Diphosphates
Egtazic Acid
Cyclic Nucleotides
Substrates
Enzymes
Chelating Agents
Substrate Specificity
Catalytic Domain
Phosphates
phosphoenolpyruvate phosphatase

Keywords

  • AphA
  • Biochemical characterization
  • class B acid phosphatase
  • Escherichia coli
  • Kinetic analysis

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Genetics

Cite this

Biochemical characterization of the class B acid phosphatase (AphA) of Escherichia coli MG1655. / Passariello, Claudio; Forleo, Costantino; Micheli, Vanna; Schippa, Serena; Leone, Rosalida; Mangani, Stefano; Thaller, Maria Cristina; Rossolini, Gian Maria.

In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1764, No. 1, 01.2006, p. 13-19.

Research output: Contribution to journalArticle

Passariello, C, Forleo, C, Micheli, V, Schippa, S, Leone, R, Mangani, S, Thaller, MC & Rossolini, GM 2006, 'Biochemical characterization of the class B acid phosphatase (AphA) of Escherichia coli MG1655', Biochimica et Biophysica Acta - Proteins and Proteomics, vol. 1764, no. 1, pp. 13-19. https://doi.org/10.1016/j.bbapap.2005.08.028
Passariello, Claudio ; Forleo, Costantino ; Micheli, Vanna ; Schippa, Serena ; Leone, Rosalida ; Mangani, Stefano ; Thaller, Maria Cristina ; Rossolini, Gian Maria. / Biochemical characterization of the class B acid phosphatase (AphA) of Escherichia coli MG1655. In: Biochimica et Biophysica Acta - Proteins and Proteomics. 2006 ; Vol. 1764, No. 1. pp. 13-19.
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