Human metastasis regulator protein H-prune is a short-chain exopolyphosphatase

Biochemistry. 2008 Sep 9;47(36):9707-13. doi: 10.1021/bi8010847. Epub 2008 Aug 14.

Abstract

The DHH superfamily human protein h-prune, a binding partner of the metastasis suppressor nm23-H1, is frequently overexpressed in metastatic cancers. From an evolutionary perspective, h-prune is very close to eukaryotic exopolyphosphatases. Here, we show for the first time that h-prune efficiently hydrolyzes short-chain polyphosphates (k cat of 3-40 s (-1)), including inorganic tripoly- and tetrapolyphosphates and nucleoside 5'-tetraphosphates. Long-chain inorganic polyphosphates (>or=25 phosphate residues) are converted more slowly, whereas pyrophosphate and nucleoside triphosphates are not hydrolyzed. The reaction requires a divalent metal cofactor, such as Mg (2+), Co (2+), or Mn (2+), which activates both the enzyme and substrate. Notably, the exopolyphosphatase activity of h-prune is suppressed by nm23-H1, long-chain polyphosphates and pyrophosphate, which may be potential physiological regulators. Nucleoside triphosphates, diadenosine hexaphosphate, cAMP, and dipyridamole (inhibitor of phosphodiesterase) do not affect this activity. Mutation of seven single residues corresponding to those found in the active site of yeast exopolyphosphatase led to a severe decrease in h-prune activity, whereas one variant enzyme exhibited enhanced activity. Our results collectively suggest that prune is the missing exopolyphosphatase in animals and support the hypothesis that the metastatic effects of h-prune are modulated by inorganic polyphosphates, which are increasingly recognized as critical regulators in cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acid Anhydride Hydrolases / genetics
  • Acid Anhydride Hydrolases / metabolism*
  • Animals
  • Coenzymes / genetics
  • Coenzymes / metabolism
  • Gene Expression Regulation, Enzymologic / genetics
  • Gene Expression Regulation, Neoplastic / genetics
  • Humans
  • Inorganic Pyrophosphatase / genetics
  • Inorganic Pyrophosphatase / metabolism
  • Metals / metabolism
  • Mutation
  • NM23 Nucleoside Diphosphate Kinases / genetics
  • NM23 Nucleoside Diphosphate Kinases / metabolism*
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism*
  • Neoplasms / enzymology*
  • Neoplasms / genetics
  • Polyphosphates / metabolism*

Substances

  • Coenzymes
  • Metals
  • NM23 Nucleoside Diphosphate Kinases
  • Neoplasm Proteins
  • Polyphosphates
  • NME1 protein, human
  • Acid Anhydride Hydrolases
  • Inorganic Pyrophosphatase
  • exopolyphosphatase