title: Structure of the kidney slit diaphragm adapter protein CD2-associated protein as determined with electron microscopy.
abstract: CD2-associated protein (CD2AP) is a multidomain scaffolding protein that has a critical role in renal function. CD2AP is expressed in glomerular podocytes at the slit diaphragm, a modified adherens junction that comprises the protein filtration barrier of the kidney, and interacts with a number of protein ligands involved in cytoskeletal remodeling, membrane trafficking, cell motility, and cell survival. The structure of CD2AP is unknown. We used electron microscopy and single particle image analysis to determine the three-dimensional structure of recombinant full-length CD2AP and found that the protein is a tetramer in solution. Image reconstruction of negatively stained protein particles generated a structure at 21 Å resolution. The protein assumed a roughly spherical, very loosely packed structure. Analysis of the electron density map revealed that CD2AP consists of a central coiled-coil domain, which forms the tetramer interface, surrounded by four symmetry-related motifs, each containing three globular domains corresponding to the three SH3 domains. The spatial organization exposes the binding sites of all 12 SH3 domains in the tetramer, allowing simultaneous binding to multiple targets. Determination of the structure of CD2AP provides novel insights into the biology of this slit diaphragm protein and lays the groundwork for characterizing the interactions between key molecules of the slit diaphragm that control glomerular filtration.
authors: Department of Medicine, Division of Nephrology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts; badair@partners.org.
authors: Brian D
authors: BD
authors: Adair
authors: Department of Medicine, Rush University Medical Center, Chicago, Illinois; and.
authors: Mehmet M
authors: MM
authors: Altintas
authors: Department of Medicine, Division of Nephrology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts;
authors: Clemens C
authors: CC
authors: Möller
authors: Department of Medicine, Division of Nephrology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts; Department of Developmental and Regenerative Biology, Harvard Medical School, Boston, Massachusetts.
authors: M Amin
authors: MA
authors: Arnaout
authors: Department of Medicine, Rush University Medical Center, Chicago, Illinois; and.
authors: Jochen
authors: J
authors: Reiser
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000648:ultrastructure*; D006801:Humans; D007678:Kidney Glomerulus; D008854:Microscopy, Electron; D050199:Podocytes; D011487:Protein Conformation
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C114545:CD2-associated protein; D003598:Cytoskeletal Proteins