A5015503966- Receptor Mechanisms and Signaling
- Estrogen and related hormone effects
- Mass Spectrometry Techniques and Applications
- Computational Drug Discovery Methods
- Metabolism, Diabetes, and Cancer
- Peroxisome Proliferator-Activated Receptors
- Adipose Tissue and Metabolism
- Immune Response and Inflammation
- Photoreceptor and optogenetics research
- Retinoids in leukemia and cellular processes
- Pancreatic function and diabetes
- Cytokine Signaling Pathways and Interactions
- Ubiquitin and proteasome pathways
- Protein Degradation and Inhibitors
- Advanced Breast Cancer Therapies
- Protein Kinase Regulation and GTPase Signaling
- Endoplasmic Reticulum Stress and Disease
- Inflammatory mediators and NSAID effects
- RNA and protein synthesis mechanisms
- Mitochondrial Function and Pathology
- Metabolomics and Mass Spectrometry Studies
- Nuclear Receptors and Signaling
- Immune cells in cancer
- Neuroscience and Neuropharmacology Research
- Advanced Data Storage Technologies
Scripps Research Institute
2014-2023
University of Florida
2022-2023
Biomedical Research Institute
2023
Universidad Cristiana Autónoma de Nicaragua
2023
Lawrence Berkeley National Laboratory
2023
Scripps (United States)
2023
The Herbert Wertheim UF Scripps Institute for Biomedical Innovation & Technology
2023
AstraZeneca (Sweden)
2021
Imperial College London
1968
Hydrogen/deuterium exchange mass spectrometry (HDX-MS) is an established method for the interrogation of protein conformation and dynamics. While data analysis challenge HDX-MS has been addressed by a number software packages, new computational tools are needed to keep pace with improved methods throughput this technique. To address these needs, we report integrated desktop program titled HDX Workbench, which facilitates automation, management, visualization, statistical cross-comparison...
Somatic mutations in the estrogen receptor alpha (ERα) gene ( ESR1 ), especially Y537S and D538G, have been linked to acquired resistance endocrine therapies. Cell-based studies demonstrated that these mutants confer ERα constitutive activity antiestrogen suggest ligand-binding domain dysfunction leads therapy resistance. Here, we integrate biophysical structural biology data reveal how lead a constitutively active antiestrogen-resistant ERα. We show mutant ERs recruit coactivator absence of...
Cereblon (CRBN) is a ubiquitin ligase (E3) substrate receptor protein co-opted by CRBN E3 modulatory drug (CELMoD) agents that target therapeutically relevant proteins for degradation. Prior crystallographic studies defined the drug-binding site within CRBN's thalidomide-binding domain (TBD), but allostery of drug-induced neosubstrate binding remains unclear. We performed cryo-electron microscopy analyses DNA damage-binding 1 (DDB1)-CRBN apo complex and compared these structures with...
The retinoic acid receptor-related orphan receptors alpha and gamma (RORalpha (NR1F1) RORgamma (NR1F3)) are nuclear perform critical roles in regulation of development, metabolism, immune function. Cholesterol cholesterol sulfate have been suggested to be RORalpha ligands, but the physiological significance is unclear. To date, no endogenous ligands described. Here, we demonstrate that 7-oxygenated sterols function as high affinity for both by directly binding their ligand-binding domains...
The nuclear receptor peroxisome proliferator-activated gamma (PPARγ) is the master regulator of adipogenesis and pharmacological target thiazolidinedione (TZD) class insulin sensitizers. Activation PPARγ by TZDs promotes at expense osteoblast formation, contributing to their associated adverse effects on bone. Recently, we reported development antagonist SR1664, designed block obesity-induced phosphorylation serine 273 (S273) in absence classical agonism, derive insulin-sensitizing efficacy...
Adenosine monophosphate (AMP)-activated protein kinase (AMPK) regulates metabolism in response to the cellular energy states. Under stress, AMP stabilizes active AMPK conformation, which activation loop (AL) is protected from phosphatases, thus keeping AL its active, phosphorylated state. At low AMP:ATP (adenosine triphosphate) ratios, ATP inhibits by increasing dynamics and accessibility. We developed conformation-specific antibodies trap ATP-bound a fully inactive, dynamic state determined...
Abstract The human mitochondrial AAA+ protein LONP1 is a critical quality control protease involved in regulating diverse aspects of biology including proteostasis, electron transport chain activity, and transcription. As such, genetic or aging-associated imbalances activity are implicated pathologic dysfunction associated with numerous diseases. Despite this importance, the molecular basis for LONP1-dependent proteolytic remains poorly defined. Here, we solved cryo-electron microscopy...
Not your typical GPCR Among the large family of G protein–coupled receptors (GPCRs) are many orphans, so called because their signaling reactions remain poorly understood. these is GPR158 which highly expressed in nervous system and implicated processes from cognition to memory mood. Patil et al . determined a high-resolution structure alone bound regulator protein (RGS) complex. has an unusual dimerization mode with extensive interaction interface that locks it conformation likely prevents...
The T cell specific RORγ isoform RORγt has been shown to be the key lineage-defining transcription factor initiate differentiation program of TH17 and TC17 cells, cells that have demonstrated antitumor efficacy. controls gene networks enhance immunity including increased IL17 production decreased immune suppression. Both synthetic putative endogenous agonists increase basal activity enhancing proliferation. Here, we show activation using drives proliferation while decreasing levels...
Abstract Circularized nandiscs (cNDs) exhibit superb monodispersity and have the potential to transform functional structural studies of membrane proteins. In particular, cNDs can stabilize large patches lipid bilayers for reconstitution complex biochemical reactions, enabling capture crucial intermediates involved in synaptic transmission viral entry. However, previous methods building require multiple steps suffer from low yields. We herein introduce a simple, one-step approach ease...
Significance To address the unmet clinical need for effectively suppressing estrogen receptor (ER) activity with both de novo resistance and in advanced ER-positive breast cancers that are resistant to standard-of-care antiestrogens, we have developed dual-mechanism ER inhibitors (DMERIs) employ two distinct ER-targeting moieties. These DMERI elicited noncanonical structural perturbations of ligand-binding domain stabilized multiple antagonist substates within dimer generate highly...
AMP-activated protein kinase (AMPK) is a central cellular energy sensor that adapts metabolism and growth to the state of cell. AMPK senses ratio adenine nucleotides (adenylate charge) by competitive binding AMP, ADP, ATP three sites (CBS1, CBS3, CBS4) in its γ-subunit. Because these are functionally interconnected, it remains unclear how bind individual sites, which occupy each site under physiological conditions, one affects other sites. Here, we comprehensively analyze nucleotide...
The vitamin D receptor/retinoid X receptor-α heterodimer (VDRRXRα) regulates bone mineralization via transcriptional control of osteocalcin (BGLAP) gene and is the receptor for 1α,25-dihydroxyvitamin D3 (1,25D3). However, supra-physiological levels 1,25D3 activates calcium-regulating TRPV6 leading to hypercalcemia. An approach attenuate this adverse effect develop selective VDR modulators (VDRMs) that differentially activate BGLAP but not TRPV6. Here we present structural insight action a...
The transcription factor estrogen receptor α (ERα) is the primary driver of ER+ breast cancer progression and a target multiple FDA-approved anticancer drugs. Ligand-dependent activity ERα determined by conformation helix-12 (H12) within ligand binding domain (LBD), but how H12 transitions from an unliganded (apo) state to active (estrogen-bound) or inactive (SERM/SERD-bound) states remains unresolved. Here, we present first crystal structure apo LBD, revealing third distinct that regulates...
The repressive states of nuclear receptors (i.e., apo or bound to antagonists inverse agonists) are poorly defined, despite the fact that a major drug target. Most ligand structures receptors, including peroxisome proliferator-activated receptor γ (PPARγ), similar structure. Here we use NMR, accelerated molecular dynamics and hydrogen-deuterium exchange mass spectrometry define PPARγ structural ensemble. We find helix 3 charge clamp positioning varies widely in is stabilized by efficacious...
Jasmonates (JAs) are plant hormones with crucial roles in development and stress resilience. They activate MYC transcription factors by mediating the proteolysis of inhibitors called JAZ proteins. In absence JA, proteins bind inhibit through assembly MYC-JAZ-Novel Interactor (NINJA)-TPL repressor complexes. However, NINJA predicted to be largely intrinsically unstructured, which has precluded their experimental structure determination. Through a combination biochemical, mutational,...
AMP-activated protein kinase (AMPK) is an attractive therapeutic target for managing metabolic diseases. A class of pharmacological activators, including Merck 991, binds the AMPK ADaM site, which forms interaction surface between domain (KD) α-subunit and carbohydrate-binding module (CBM) β-subunit. Here, we report development two new 991-derivative compounds, R734 R739, potently activate in a variety cell types, β2-specific skeletal muscle cells. Surprisingly, found that they have only...
Retinoic acid inducible gene-I (RIG-I) ensures immune surveillance of viral RNAs bearing a 5'-triphosphate (5'ppp) moiety. Mutations in RIG-I (C268F and E373A) lead to impaired ATPase activity, thereby driving hyperactive signaling associated with autoimmune diseases. Here we report, using hydrogen/deuterium exchange, mechanistic models for dysregulated proofreading that ultimately result the improper recognition cellular 7-methylguanosine N1-2'-O-methylation (Cap1) on 5' end. Cap1-RNA...