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

doi:10.1074/jbc.ra119.008218

Title: Dynamic structure of the full-length scaffolding protein NHERF1 influences signaling complex assembly

Journal Article · 06 June 2019 · Journal of Biological Chemistry

DOI: https://doi.org/10.1074/jbc.ra119.008218 · OSTI ID:1545195

Bhattacharya, Shibani ^[1];

^[2];

^[3]; Bu, Zimei ^[4]

New York Structural Biology Center, NY (United States)
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Univ. of Pittsburgh, PA (United States)
City Univ. (CUNY), NY (United States)

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. We applied in this report 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). Significant insight into the modulation of the intrinsic flexibility of NHERF1 by disease-associated point mutations that alter the dynamic assembly of signaling complexes are provided in this work.

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Research Organization:: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22), National Institutes of Health (NIH); National Science Foundation (NSF); U.S. Dept. of Defense

Grant/Contract Number:: AC05-00OR22725

OSTI ID:: 1545195

Journal Information:: Journal of Biological Chemistry, Journal Name: Journal of Biological Chemistry Journal Issue: 29 Vol. 294; ISSN 0021-9258

Publisher:: American Society for Biochemistry and Molecular BiologyCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
G protein-coupled receptor kinase 6A (GRK6A)
NHERF1
NMR
PDZ domain
SLC9A3 regulator 1 (SLC9A3R1)
intracellular transport
intrinsically disordered protein
scaffolding protein
small-angle neutron scattering (SANS)

Title: Dynamic structure of the full-length scaffolding protein NHERF1 influences signaling complex assembly

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