Structural and enzymatic characterization of the streptococcal ATP/diadenosine polyphosphate and phosphodiester hydrolase Spr1479/SapH

J Biol Chem. 2011 Oct 14;286(41):35906-35914. doi: 10.1074/jbc.M111.228585. Epub 2011 Aug 23.

Abstract

Spr1479 from Streptococcus pneumoniae R6 is a 33-kDa hypothetical protein of unknown function. Here, we determined the crystal structures of its apo-form at 1.90 Å and complex forms with inorganic phosphate and AMP at 2.30 and 2.20 Å, respectively. The core structure of Spr1479 adopts a four-layer αββα-sandwich fold, with Fe(3+) and Mn(2+) coordinated at the binuclear center of the active site (similar to metallophosphoesterases). Enzymatic assays showed that, in addition to phosphodiesterase activity for bis(p-nitrophenyl) phosphate, Spr1479 has hydrolase activity for diadenosine polyphosphate (Ap(n)A) and ATP. Residues that coordinate with the two metals are indispensable for both activities. By contrast, the streptococcus-specific residue Trp-67, which binds to phosphate in the two complex structures, is indispensable for the ATP/Ap(n)A hydrolase activity only. Moreover, the AMP-binding pocket is conserved exclusively in all streptococci. Therefore, we named the protein SapH for streptococcal ATP/Ap(n)A and phosphodiester hydrolase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acid Anhydride Hydrolases / chemistry*
  • Adenosine Triphosphatases / chemistry*
  • Apoenzymes / chemistry
  • Bacterial Proteins / chemistry*
  • Binding Sites
  • Crystallography, X-Ray
  • Protein Folding*
  • Protein Structure, Secondary
  • Streptococcus pneumoniae / enzymology*

Substances

  • Apoenzymes
  • Bacterial Proteins
  • Acid Anhydride Hydrolases
  • Adenosine Triphosphatases
  • diadenosine polyphosphate hydrolase

Associated data

  • PDB/3QFM
  • PDB/3QFN
  • PDB/3QFO