Causal link between nucleotide pyrophosphohydrolase overactivity and increased intracellular inorganic pyrophosphate generation demonstrated by transfection of cultured fibroblasts and osteoblasts with plasma cell membrane glycoprotein-1. Relevance to calcium pyrophosphate dihydrate deposition disease.
OBJECTIVE
In subjects with idiopathic calcium pyrophosphate dihydrate (CPPD) deposition disease, cartilage chondrocytes elaborate increased amounts of PPi. The mechanism of the intracellular PPi elevation is not known. Plasma membrane 5'-nucleotide phosphodiesterase I/nucleotide pyrophosphohydrolase (NTPPPH) activity also is elevated in chondrocytes and dermal fibroblasts of patients with idiopathic CPPD deposition disease. NTPPPH, as an ecto-enzyme, could act within certain intracellular compartments. Thus, we hypothesized a potential causal link between increased NTPPPH activity and increased intracellular PPi.


METHODS
Transformed simian fibroblasts (COS cells) and human osteoblasts (U2OS cells) were transfected with the 5'-nucleotide phosphodiesterase I ecto-enzyme plasma cell membrane glycoprotein-1 (PC-1), recently shown to be expressed in cartilage, osteoblasts, and fibroblasts.


RESULTS
Transfection with PC-1 markedly up-regulated 5'-nucleotode phosphodiesterase I activity and increased intracellular PPi concentrations by increasing the capacity of cells to generate PPi. Importantly, this did not require supplementation with exogenous nucleotides.


CONCLUSIONS
Cellular overexpression of PC-1 produces NTPPPH overactivity and increased intracellular PPi generation in vitro. These findings support the potential importance of NTPPPH overactivity in PPi generation, both inside and outside the cell, in some subjects with CPPD deposition disease.
Terkeltaub
R
R
Veterans Affairs Medical Center, University of California San Diego.
Rosenbach
M
M
Fong
F
F
Goding
J
J
D000818:Animals; D002128:Calcium Metabolism Disorders / Q000201:enzymology / Q000378:metabolism; D002478:Cells, Cultured; D011756:Diphosphates / Q000378:metabolism*; D005347:Fibroblasts / Q000201:enzymology / Q000378:metabolism; D000882:Haplorhini; D006801:Humans; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000494:pharmacology*; D010006:Osteoblasts / Q000201:enzymology / Q000378:metabolism; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases / Q000187:drug effects / Q000378:metabolism*; D011755:Pyrophosphatases / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D014162:Transfection
D011756:Diphosphates; D008562:Membrane Glycoproteins; D010727:Phosphoric Diester Hydrolases; D043264:Phosphodiesterase I; C065962:ectonucleotide pyrophosphatase phosphodiesterase 1; D011755:Pyrophosphatases; C021895:nucleoside triphosphate pyrophosphatase