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The CPPDD-associated ANKH M48T mutation interrupts the interaction of ANKH with the sodium/phosphate cotransporter

John Wang1, Hing Wo Tsui, Frank Beier

  • 1Toronto Western Research Institute, University Health Network, University of Toronto, and Toronto Western Hospital, Mc14-419, 399 Bathurst Street, Toronto, Ontario M5T 2S8, Canada.

The Journal of Rheumatology
|April 17, 2009
PubMed

Insights

Mutations in the progressive ankylosis (ANKH) gene are linked to calcium pyrophosphate dihydrate crystal deposition disease (CPPDD). This study reveals ANKH interacts with PiT-1, and a CPPDD-associated mutation disrupts this interaction.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genetics

Background:

  • Familial calcium pyrophosphate dihydrate crystal deposition disease (CPPDD) is often associated with dominant mutations in the human homolog of progressive ankylosis (ANKH) gene.
  • The dominant inheritance pattern of these mutations suggests ANKH may interact with other proteins.

Purpose of the Study:

  • To investigate protein interactions of ANKH.
  • To determine if CPPDD-associated ANKH mutations disrupt these interactions.

Main Methods:

  • Generated stable ATDC5 cells expressing wild-type (wt) ANKH and the ANKH M48T mutant.
  • Used coimmunoprecipitation and Western blot to identify ANKH-interacting proteins.
  • Assessed the impact of high phosphate on gene expression related to phosphate (Pi) and pyrophosphate (PPi) homeostasis.

Main Results:

  • Demonstrated that ANKH protein physically associates with the sodium/phosphate cotransporter PiT-1.
  • Showed that the ANKH M48T mutant protein is unable to interact with PiT-1.
  • Observed that high phosphate treatment disrupted the coordinated upregulation of Ank and PiT1 transcripts in ANKH M48T cells.

Conclusions:

  • Established a coordinated interrelationship between ANKH and PiT-1 in Pi and PPi metabolism.
  • Suggests that disruption of the ANKH-PiT-1 interaction by CPPDD-associated mutations may contribute to disease pathogenesis.
Abstract

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