A5038436171- Parathyroid Disorders and Treatments
- Metabolism, Diabetes, and Cancer
- Protein Structure and Dynamics
- Ion Transport and Channel Regulation
- Genetic Syndromes and Imprinting
- Protein Kinase Regulation and GTPase Signaling
- Magnesium in Health and Disease
- RNA and protein synthesis mechanisms
- Hippo pathway signaling and YAP/TAZ
- Erythrocyte Function and Pathophysiology
- Receptor Mechanisms and Signaling
- Eastern European Communism and Reforms
- Biomedical Research and Pathophysiology
- Chemical Synthesis and Analysis
- Neuroscience and Neuropharmacology Research
- Soviet and Russian History
- Crystallography and molecular interactions
- Enzyme Structure and Function
- Mechanisms of cancer metastasis
- Computational Drug Discovery Methods
- Hedgehog Signaling Pathway Studies
- Nitric Oxide and Endothelin Effects
- Fibroblast Growth Factor Research
- Neurofibromatosis and Schwannoma Cases
- Amino Acid Enzymes and Metabolism
University of Pittsburgh
2012-2024
Carnegie Mellon University
2005-2012
Proteins are held together in the native state by hydrophobic interactions, hydrogen bonds and interactions with surrounding water, whose strength as well spatial temporal distribution affects protein flexibility hence function. We study these effects using 10 ns molecular dynamics simulations of pure water two proteins, glutamate receptor ligand binding domain barnase. find that most noncovalent flicker on off over typically nanoseconds, so we can obtain good statistics from simulations....
The Na(+)/H(+) exchange regulatory factor-1 (NHERF1) is a scaffolding protein that possesses two tandem PDZ domains and carboxy-terminal ezrin-binding domain (EBD). parathyroid hormone receptor (PTHR), type II sodium-dependent phosphate cotransporter (Npt2a), β2-adrenergic (β2-AR), through their respective PDZ-recognition motifs, individually interact with NHERF1 forming complex one of the domains. In basal state, adopts self-inhibited conformation, in which its ligand interacts PDZ2. We...
Significance Our findings identify a previously unidentified role for scaffolding protein ezrin-radixin-moesin-binding phosphoprotein 50 (EBP50; aka NHERF1) in the activation of NADPH oxidases (Nox), family professional reactive oxygen species (ROS) producing enzymes implicated numerous pathologies. We demonstrate that EBP50 is critical agonist-induced production ROS superoxide anion (O 2 •– ), and it directly associates with Nox organizing subunit p47 phox . deletion abolishes angiotensin...
SARS-CoV-2 is responsible for the global COVID-19 pandemic. Angiotensin converting enzyme 2 (ACE2) membrane-delimited receptor SARS-CoV-2. Lung, intestine, and kidney, major sites of viral infection, express ACE2 that harbors an intracellular, carboxy-terminal PDZ-recognition motif. These organs prominently PDZ protein Na+/H+ exchanger regulatory factor-1 (NHERF1). Here, we report NHERF1 tethers augments cell entry. directly binds both domains. Disruption either core-binding motif or...
Phosphate homeostasis, mediated by dietary intake, renal absorption, and bone deposition, is incompletely understood because of the uncharacterized roles numerous implicated protein factors. Here, we identified a novel role for one such element, regulator G signaling 14 (RGS14), suggested genome-wide association studies to associate with dysregulated Pi levels. We show that human RGS14 possesses carboxy-terminal PDZ ligand required sodium phosphate cotransporter 2a (NPT2A) hydrogen exchanger...
Na+/H+ Exchanger Regulatory Factor-1 (NHERF1) is a scaffolding protein containing 2 PDZ domains that coordinates the assembly and trafficking of transmembrane receptors ion channels. Most target proteins harboring C-terminus recognition motif bind more-or-less equivalently to either domain, which contain identical core-binding motifs. However some substrates such as type II sodium-dependent phosphate co-transporter (NPT2A), uniquely only one domain. We sought define structural determinants...
The ionotropic glutamate receptors are localized in the pre- and postsynaptic membrane of neurons brain. Activation by principal excitatory neurotransmitter allows ligand binding domain to change conformation, communicating opening channel for ion conduction. free energy GluR2 S1S2 (S1S2) closure transition was computed using a combination thermodynamic integration umbrella sampling modeling methods. A path that involves lowering charge on E705 chosen clarify role this site residue....
Na+/H+ exchanger regulatory factor-1 (NHERF1) is a scaffolding protein containing two PSD95/discs large protein/ZO1 (PDZ) domains that modifies the signaling, trafficking, and function of parathyroid hormone receptor (PTHR), family B G-protein-coupled receptor. PTHR NHERF1 bind through PDZ–ligand-recognition mechanism. We show PTH elicits phosphorylation Thr591 in canonical -ETVM binding motif PTHR. Conservative substitution with Cys does not affect PTH(1–34)-induced cAMP production or to...
Abstract Parathyroid hormone (PTH) and fibroblast growth factor-23 (FGF23) control extracellular phosphate levels by regulating renal NPT2A-mediated transport a process requiring the PDZ scaffold protein NHERF1. NHERF1 possesses two domains, PDZ1 PDZ2, with identical core-binding GYGF motifs explicitly recognizing distinct binding partners that play different specific roles in hormone-regulated transport. The interaction of carboxy-terminal PDZ-binding motif NPT2A (C-TRL) is required for...
Hydrogen–deuterium exchange-mass spectrometry (HDXMS) is a powerful technology to characterize conformations and conformational dynamics of proteins protein complexes. HDXMS has been widely used in the field therapeutics for development drugs. Although sufficient sequence coverage critical success HDXMS, it sometimes difficult achieve. In this study, we developed data analysis strategy that includes parallel post-translational modification (PTM) scanning analysis. Using membrane-delimited G...
The type II sodium-dependent phosphate cotransporter (NPT2A) mediates renal uptake. NPT2A is regulated by parathyroid hormone (PTH) and fibroblast growth factor 23, which requires Na+/H+ exchange regulatory factor-1 (NHERF1), a multidomain PDZ-containing phosphoprotein. Phosphocycling controls the association between NHERF1 NPT2A. Here, we characterize critical involvement of G protein–coupled receptor kinase 6A (GRK6A) in mediating PTH-sensitive transport targeted phosphorylation coupled...
Using molecular dynamics (MD) simulations, computational protein modifications, and a novel theoretical methodology that determines structural rigidity/flexibility (the FIRST algorithm), we investigate how structure of the glutamate receptor ligand binding domain (GluR2 S1S2) facilitate its conformational transition. S1S2 is two-lobe protein, which undergoes cleft closure transition upon an agonist in between two lobes; hence it expected mechanism this can be characterized as hinge-type....
To function properly protein molecules require both flexibility and rigidity, therefore fast accurate prediction of rigidity/flexibility is one the important problems in science. In this work we used two theoretical approaches to determine flexible regions four homologous pairs proteins from thermophilic mesophilic organisms. Protein chosen study were selected represent typical folding classes. Our first approach, FoldUnfold, uses amino acid sequence statistical information on density...
The Na+-dependent phosphate cotransporter-2A (NPT2A, SLC34A1) is a primary regulator of extracellular homeostasis. Its most prominent structural element carboxy-terminal PDZ ligand that binds Na+/H+ Exchanger Regulatory Factor-1 (NHERF1, SLC9A3R1). NHERF1, multidomain protein, establishes NPT2A membrane localization and required for hormone-inhibitable transport. also possesses an uncharacterized internal ligand. Two recent clinical reports describe congenital hypophosphatemia in children...
The ion channel protein α-hemolysin (αHL) forms supramolecular complexes with the polysaccharide β-cyclodextrin (βCD). This system has potential uses in nanoscale device engineering. It been found recently that βCD formed longer- or shorter-lived some engineered αHL mutants then a wild type (Gu et al. J. Gen. Physiol. 2001, 118, 481−493). However, how changes sequence affect complex lifetime was not completely understood part due to lack of knowledge structures these metastable complexes. In...
RGS14 is a multifunctional scaffolding protein that integrates G protein, MAPK, and Ca ++ /CaM signaling pathways. contains an RGS domain, tandem Ras/Rap-binding domains, regulatory motif. Human/primate differ from rodent proteins by possessing carboxy-terminal extension encoding Type I PDZ ligand. has been implicated in disordered phosphate metabolism. The human gene adjacent to SLC34A1 encodes the NPT2A sodium-phosphate cotransporter. Hormone-regulated requires NHERF1 which two domains...
G protein-coupled receptors, including PTHR, are pivotal for controlling metabolic processes ranging from serum phosphate and vitamin D levels to glucose uptake, cytoplasmic interactors may modulate their signaling, trafficking, function. We now show that direct interaction with Scribble, a cell polarity-regulating adaptor protein, modulates PTHR activity. Scribble is crucial regulator establishing developing tissue architecture, its dysregulation involved in various disease conditions,...