Ezrin induces long-range interdomain allostery in the scaffolding protein NHERF1.
Scaffolding proteins are molecular switches that control diverse signaling events. The scaffolding protein Na(+)/H(+) exchanger regulatory factor 1 (NHERF1) assembles macromolecular signaling complexes and regulates the macromolecular assembly, localization, and intracellular trafficking of a number of membrane ion transport proteins, receptors, and adhesion/antiadhesion proteins. NHERF1 begins with two modular protein-protein interaction domains-PDZ1 and PDZ2-and ends with a C-terminal (CT) domain. This CT domain binds to ezrin, which, in turn, interacts with cytosekeletal actin. Remarkably, ezrin binding to NHERF1 increases the binding capabilities of both PDZ domains. Here, we use deuterium labeling and contrast variation neutron-scattering experiments to determine the conformational changes in NHERF1 when it forms a complex with ezrin. Upon binding to ezrin, NHERF1 undergoes significant conformational changes in the region linking PDZ2 and its CT ezrin-binding domain, as well as in the region linking PDZ1 and PDZ2, involving very long range interactions over 120 A. The results provide a structural explanation, at mesoscopic scales, of the allosteric control of NHERF1 by ezrin as it assembles protein complexes. Because of the essential roles of NHERF1 and ezrin in intracellular trafficking in epithelial cells, we hypothesize that this long-range allosteric regulation of NHERF1 by ezrin enables the membrane-cytoskeleton to assemble protein complexes that control cross-talk and regulate the strength and duration of signaling.
Li
Jianquan
J
Fox Chase Cancer Center, Philadelphia, PA 19111, USA.
Callaway
David J E
DJ
Bu
Zimei
Z
D000494:Allosteric Regulation; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D003903:Deuterium / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008954:Models, Biological; D008958:Models, Molecular; D010750:Phosphoproteins / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D053838:Scattering, Small Angle; D017923:Sodium-Hydrogen Exchangers / Q000737:chemistry* / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods; D000099149:Ezrin
D003598:Cytoskeletal Proteins; D003903:Deuterium; D010750:Phosphoproteins; D017923:Sodium-Hydrogen Exchangers; D000099149:Ezrin; C093376:sodium-hydrogen exchanger regulatory factor