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. © 2005 Elsevier B.V. All rights reserved.

Biochemical characterization of the class B acid phosphatase (AphA) of Escherichia coli MG1655 / Passariello, Claudio; Costantino, Forleo; Vanna, Micheli; Schippa, Serena; Rosalida, Leone; Stefano, Mangani; Maria Cristina, Thaller; Gian Maria, Rossolini. - In: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS. - ISSN 1570-9639. - 1764:1(2006), pp. 13-19. [10.1016/j.bbapap.2005.08.028]

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

PASSARIELLO, Claudio;SCHIPPA, Serena;
2006

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. © 2005 Elsevier B.V. All rights reserved.
2006
apha; biochemical characterization; class b acid phosphatase; escherichia coli; kinetic analysis
01 Pubblicazione su rivista::01a Articolo in rivista
Biochemical characterization of the class B acid phosphatase (AphA) of Escherichia coli MG1655 / Passariello, Claudio; Costantino, Forleo; Vanna, Micheli; Schippa, Serena; Rosalida, Leone; Stefano, Mangani; Maria Cristina, Thaller; Gian Maria, Rossolini. - In: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS. - ISSN 1570-9639. - 1764:1(2006), pp. 13-19. [10.1016/j.bbapap.2005.08.028]
File allegati a questo prodotto
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/122604
 Attenzione

Attenzione! I dati visualizzati non sono stati sottoposti a validazione da parte dell'ateneo

Citazioni
  • ???jsp.display-item.citation.pmc??? 12
  • Scopus 20
  • ???jsp.display-item.citation.isi??? 20
social impact