Your browser doesn't support javascript.
loading
Dynamic structure of the full-length scaffolding protein NHERF1 influences signaling complex assembly.
Bhattacharya, Shibani; Stanley, Christopher B; Heller, William T; Friedman, Peter A; Bu, Zimei.
Afiliación
  • Bhattacharya S; New York Structural Biology Center, New York, New York 10027 sbhattacharya@nysbc.org.
  • Stanley CB; Neutron Scattering Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37830.
  • Heller WT; Neutron Scattering Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37830.
  • Friedman PA; Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, Pennsylvania 15261 paf10@pitt.edu.
  • Bu Z; Department of Chemistry and Biochemistry, City College of New York, New York, New York 10031 zbu@ccny.cuny.edu.
J Biol Chem ; 294(29): 11297-11310, 2019 07 19.
Article en En | MEDLINE | ID: mdl-31171716
The Na+/H+ exchange regulatory cofactor 1 (NHERF1) protein modulates the assembly and intracellular trafficking of several transmembrane G protein-coupled receptors (GPCRs) and ion transport proteins with the membrane-cytoskeleton adapter protein ezrin. Here, we applied solution NMR and small-angle neutron scattering (SANS) to structurally characterize full-length NHERF1 and disease-associated variants that are implicated in impaired phosphate homeostasis. Using NMR, we mapped the modular architecture of NHERF1, which is composed of two structurally-independent PDZ domains that are connected by a flexible, disordered linker. We observed that the ultra-long and disordered C-terminal tail of NHERF1 has a type 1 PDZ-binding motif that interacts weakly with the proximal, second PDZ domain to form a dynamically autoinhibited structure. Using ensemble-optimized analysis of SANS data, we extracted the molecular size distribution of structures from the extensive conformational space sampled by the flexible chain. Our results revealed that NHERF1 is a diffuse ensemble of variable PDZ domain configurations and a disordered C-terminal tail. The joint NMR/SANS data analyses of three disease variants (L110V, R153Q, and E225K) revealed significant differences in the local PDZ domain structures and in the global conformations compared with the WT protein. Furthermore, we show that the substitutions affect the affinity and kinetics of NHERF1 binding to ezrin and to a C-terminal peptide from G protein-coupled receptor kinase 6A (GRK6A). These findings provide important insight into the modulation of the intrinsic flexibility of NHERF1 by disease-associated point mutations that alter the dynamic assembly of signaling complexes.
Asunto(s)
Palabras clave
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Fosfoproteínas / Transducción de Señal / Intercambiadores de Sodio-Hidrógeno Límite: Humans Idioma: En Revista: J Biol Chem Año: 2019 Tipo del documento: Article Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Fosfoproteínas / Transducción de Señal / Intercambiadores de Sodio-Hidrógeno Límite: Humans Idioma: En Revista: J Biol Chem Año: 2019 Tipo del documento: Article Pais de publicación: Estados Unidos