A high-resolution structure of the EF-hand domain of human polycystin-2

Protein Sci. 2014 Sep;23(9):1301-8. doi: 10.1002/pro.2513. Epub 2014 Jul 22.

Abstract

Autosomal dominant polycystic kidney disease (ADPKD) affects over 1:1000 of the worldwide population and is caused by mutations in two genes, PKD1 and PKD2. PKD2 encodes a 968-amino acid membrane spanning protein, Polycystin-2 (PC-2), which is a member of the TRP ion channel family. The C-terminal cytoplasmic tail contains an EF-hand motif followed by a short coiled-coil domain. We have determined the structure of the EF-hand region of PC-2 using NMR spectroscopy. The use of different boundaries, compared with those used in previous studies, have enabled us to determine a high resolution structure and show that the EF hand motif forms a standard calcium-binding pocket. The affinity of this pocket for calcium has been measured and mutants that both decrease and increase its affinity for the metal ion have been created.

Keywords: ADPKD; EF-hand; ITC; NMR; mutagenesis; polycystin-2; solution structure.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcium / chemistry
  • Calcium / metabolism
  • Calorimetry
  • EF Hand Motifs*
  • Humans
  • Models, Molecular
  • Nuclear Magnetic Resonance, Biomolecular
  • Protein Conformation
  • TRPP Cation Channels / chemistry*
  • TRPP Cation Channels / genetics
  • TRPP Cation Channels / metabolism

Substances

  • TRPP Cation Channels
  • polycystic kidney disease 2 protein
  • Calcium