Concerted regulation of inorganic pyrophosphate and osteopontin by akp2, enpp1, and ank: an integrated model of the pathogenesis of mineralization disorders

Am J Pathol. 2004 Apr;164(4):1199-209. doi: 10.1016/S0002-9440(10)63208-7.


Tissue-nonspecific alkaline phosphatase (TNAP) hydrolyzes the mineralization inhibitor inorganic pyrophosphate (PP(i)). Deletion of the TNAP gene (Akp2) in mice results in hypophosphatasia characterized by elevated levels of PP(i) and poorly mineralized bones, which are rescued by deletion of nucleotide pyrophosphatase phosphodiesterase 1 (NPP1) that generates PP(i). Mice deficient in NPP1 (Enpp1(-/-)), or defective in the PP(i) channeling function of ANK (ank/ank), have decreased levels of extracellular PP(i) and are hypermineralized. Given the similarity in function between ANK and NPP1 we crossbred Akp2(-/-) mice to ank/ank mice and found a partial normalization of the mineralization phenotypes and PP(i) levels. Examination of Enpp1(-/-) and ank/ank mice revealed that Enpp1(-/-) mice have a more severe hypermineralized phenotype than ank/ank mice and that NPP1 but not ANK localizes to matrix vesicles, suggesting that failure of ANK deficiency to correct hypomineralization in Akp2(-/-) mice reflects the lack of ANK activity in the matrix vesicle compartment. We also found that the mineralization inhibitor osteopontin (OPN) was increased in Akp2(-/-), and decreased in ank/ank mice. PP(i) and OPN levels were normalized in [Akp2(-/-); Enpp1(-/-)] and [Akp2(-/-); ank/ank] mice, at both the mRNA level and in serum. Wild-type osteoblasts treated with PP(i) showed an increase in OPN, and a decrease in Enpp1 and Ank expression. Thus TNAP, NPP1, and ANK coordinately regulate PP(i) and OPN levels. The hypomineralization observed in Akp2(-/-) mice arises from the combined inhibitory effects of PP(i) and OPN. In contrast, NPP1 or ANK deficiencies cause a decrease in the PP(i) and OPN pools that leads to hypermineralization.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alkaline Phosphatase / physiology*
  • Animals
  • Animals, Genetically Modified
  • Blotting, Western
  • Bone and Bones / physiology
  • Calcification, Physiologic / physiology*
  • Cells, Cultured
  • Diphosphates / metabolism*
  • Diphosphates / pharmacology
  • Enzyme-Linked Immunosorbent Assay
  • Inorganic Chemicals
  • Membrane Proteins / deficiency
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Mice
  • Models, Biological
  • Osteoblasts / drug effects
  • Osteopontin
  • Phosphate Transport Proteins
  • Phosphoric Diester Hydrolases / deficiency
  • Phosphoric Diester Hydrolases / genetics
  • Phosphoric Diester Hydrolases / metabolism
  • Pyrophosphatases / deficiency
  • Pyrophosphatases / genetics
  • Pyrophosphatases / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sialoglycoproteins / metabolism*


  • Diphosphates
  • Inorganic Chemicals
  • Membrane Proteins
  • Phosphate Transport Proteins
  • Sialoglycoproteins
  • Spp1 protein, mouse
  • ank protein, mouse
  • Osteopontin
  • Alkaline Phosphatase
  • Phosphoric Diester Hydrolases
  • ectonucleotide pyrophosphatase phosphodiesterase 1
  • Pyrophosphatases