title: A high-resolution structure of the EF-hand domain of human polycystin-2.
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.
authors.affiliation: MRC Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 0QH, United Kingdom.
authors.forename: Mark D
authors.initials: MD
authors.lastname: Allen
authors.forename: Seema
authors.initials: S
authors.lastname: Qamar
authors.forename: Murali K
authors.initials: MK
authors.lastname: Vadivelu
authors.forename: Richard N
authors.initials: RN
authors.lastname: Sandford
authors.forename: Mark
authors.initials: M
authors.lastname: Bycroft
mesh_terms: D002118:Calcium / Q000737:chemistry / Q000378:metabolism; D002151:Calorimetry; D020832:EF Hand Motifs; D006801:Humans; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D050396:TRPP Cation Channels / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D002118:Calcium; D050396:TRPP Cation Channels
keywords: ADPKD; EF-hand; ITC; NMR; mutagenesis; polycystin-2; solution structure