title: Atomic force microscopy imaging reveals the domain structure of polycystin-1.
abstract: Mutation of polycystin-1 (PC1) is the major cause of autosomal dominant polycystic kidney disease. PC1 has a predicted molecular mass of ~460 kDa comprising a long multidomain extracellular N-terminal region, 11 transmembrane regions, and a short C-terminal region. Because of its size, PC1 has proven difficult to handle biochemically, and structural information is consequently sparse. Here we have isolated wild-type PC1, and several mutants, from transfected cells by immunoaffinity chromatography and visualized individual molecules using atomic force microscopy (AFM) imaging. Full-length PC1 appeared as two unequally sized blobs connected by a 35 nm string. The relative sizes of the two blobs suggested that the smaller one represents the N-terminus, including the leucine-rich repeats, the first polycystic kidney disease (PKD) domain, and the C-type lectin motif, while the larger one is the C-terminus, including the receptor for egg jelly (REJ) domain, all transmembrane domains, and the cytoplasmic tail. The intervening string would then consist of a series of tandem PKD domains. The structures of the various PC1 mutants were all consistent with this model. Our results represent the first direct visualization of the structure of PC1, and reveal the architecture of the protein, with intriguing implications for its function.
authors.affiliation: Department of Pharmacology, University of Cambridge, Cambridge CB2 1PD, UK.
authors.forename: Peter
authors.initials: P
authors.lastname: Oatley
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Stewart
authors.forename: Richard
authors.initials: R
authors.lastname: Sandford
authors.forename: J Michael
authors.initials: JM
authors.lastname: Edwardson
mesh_terms: D006801:Humans; D018625:Microscopy, Atomic Force / Q000379:methods*; D011487:Protein Conformation; D050396:TRPP Cation Channels / Q000737:chemistry*
chemical_list: D050396:TRPP Cation Channels