The molecular class C acid phosphatase of Chryseobacterium meningosepticum (OlpA) is a broad-spectrum nucleotidase with preferential activity on 5′-nucleotides

Claudio Passariello, Serena Schippa, Patrizia Iori, Francesca Berlutti, Maria Cristina Thaller, Gian Maria Rossolini

Research output: Contribution to journalArticle

Abstract

The olpA gene of Chryseobacterium meningosepticum, encoding a molecular class C phosphatase, was cloned and expressed in Escherichia coli. The gene encodes a 29-kDa polypeptide containing an amino-terminal signal peptide typical of bacterial membrane lipoproteins. Expression in E. coli results in a functional product that mostly partitions in the outer membrane. A secreted soluble OlpA derivative (sOlpA) lacking the N-terminal cysteine residue for lipid anchoring was produced in E. coli and purified by means of two steps of ion exchange chromatography. Analysis of the kinetic parameters of sOlpA with several organic phosphoesters revealed that the enzyme was able to efficiently hydrolyze nucleotide monophosphates, with a strong preference for 5′-nucleotides and for 3′-AMP. The enzyme was also able to hydrolyze sugar phosphates and β-glycerol phosphate, although with a lower efficiency, whereas it was apparently inactive against nucleotide di- and triphosphates, diesters, and phytate. OlpA, therefore, can be considered a broad-spectrum nucleotidase with preference for 5′-nucleotides. Its functional behaviour exhibits differences from that of the Haemophilus influenzae OMP P4 lipoprotein, revealing functional heterogeneity among phosphatases of molecular class C.

Original languageEnglish
Pages (from-to)203-209
Number of pages7
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1648
Issue number1-2
DOIs
Publication statusPublished - May 30 2003

Keywords

  • Acid phosphatase
  • Chryseobacterium meningosepticum
  • Nucleotidase

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Genetics

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