A5011003940- Receptor Mechanisms and Signaling
- Monoclonal and Polyclonal Antibodies Research
- Neuropeptides and Animal Physiology
- Cytokine Signaling Pathways and Interactions
- Chronic Lymphocytic Leukemia Research
- Chemokine receptors and signaling
- HER2/EGFR in Cancer Research
- Signaling Pathways in Disease
- RNA Interference and Gene Delivery
- Advanced Biosensing Techniques and Applications
- Pharmacological Receptor Mechanisms and Effects
- Axon Guidance and Neuronal Signaling
- RNA and protein synthesis mechanisms
- Single-cell and spatial transcriptomics
- Computational Drug Discovery Methods
- Viral Infectious Diseases and Gene Expression in Insects
- Advanced biosensing and bioanalysis techniques
- T-cell and B-cell Immunology
- Mycobacterium research and diagnosis
- Renin-Angiotensin System Studies
- Lymphoma Diagnosis and Treatment
- Protein purification and stability
- Myeloproliferative Neoplasms: Diagnosis and Treatment
- Biosensors and Analytical Detection
- Phosphodiesterase function and regulation
Primity Bio (United States)
2014-2020
DiscoveRx (United States)
2009-2018
IDEX Corporation (United States)
2013
Stanford University
2002-2007
Baxter (United States)
2002-2007
We have defined inactive α and ω fragments of β-lactamase that can complement to form a functional enzyme in both bacteria mammalian cells, serving as readout for the interaction proteins fused fragments. Critical this advance was identification tripeptide, Asn-Gly-Arg, which when juxtaposed at carboxyl terminus fragment increased complemented activity by up 4 orders magnitude. β-Lactamase is well suited monitoring constitutive inducible protein interactions because it small (29 kDa),...
A variety of G-protein–coupled receptor (GPCR) screening technologies have successfully partnered a number GPCRs with their cognate ligands. GPCR-mediated β-arrestin recruitment is now recognized as distinct intracellular signaling pathway, and ligand-receptor interactions may show bias toward over classical GPCR pathways. We hypothesized that the failure to identify native ligands for remaining orphan be consequence biased signaling. To investigate this, we assembled 10 500 candidate...
μ-Opioid receptors are among the most studied G protein-coupled because of therapeutic value agonists, such as morphine, that used to treat chronic pain. However, these drugs have significant side effects, respiratory suppression, constipation, allodynia, tolerance, and dependence, well abuse potential. Efforts fine tune pain control while alleviating effects drugs, both physiological psychological, led development a wide variety structurally diverse agonist ligands for μ-opioid receptor,...
Chemokines display considerable promiscuity with multiple ligands and receptors shared in common, a phenomenon that is thought to underlie their biochemical "redundancy." Their are part of larger seven-transmembrane receptor superfamily, commonly referred as G protein-coupled receptors, which have been demonstrated be able signal different efficacies downstream signaling pathways, biased agonism. Biased agonism has primarily reported synthetic ligands, the biologic prevalence importance such...
Most secreted growth factors and cytokines are functionally pleiotropic because their receptors expressed on diverse cell types. While important for normal mammalian physiology, pleiotropy limits the efficacy of as therapeutics. Stem factor (SCF) is a that acts through c-Kit receptor tyrosine kinase to elicit hematopoietic progenitor expansion but can be toxic when administered in vivo it concurrently activates mast cells. We engineered mechanism-based SCF partial agonist impaired...
Allosteric modulators of G protein-coupled receptors (GPCRs) have a number potential advantages compared to agonists or antagonists that bind the orthosteric site receptor. These include for receptor selectivity, maintenance temporal and spatial fidelity signaling in vivo, ceiling effect allosteric cooperativity which may prevent overdose issues, engendering bias by differentially modulating distinct pathways. Here we describe discovery, synthesis, molecular pharmacology δ-opioid...
Cytokine and growth-factor ligands typically signal through homo- or hetero-dimeric cell surface receptors via Janus Kinase (JAK/TYK), Receptor Tyrosine (RTK)-mediated trans-phosphorylation. However, the number of receptor dimer pairings occurring in nature is limited to those driven by natural encoded within our genome. We have engineered synthethic cytokines (synthekines) that drive formation cytokine are not formed endogenous found nature, which activate distinct signaling programs. show...
The orphan receptor tyrosine kinase ErbB2 is activated by each of the EGFR family members upon ligand binding. However, difficulties monitoring dynamic interactions membrane receptors have hindered elucidation mechanism activation. We engineered a system to monitor protein-protein in intact mammalian cells such that different sets protein can be quantitatively compared. Application this showed interacts stably with and ErbB3, but fails spontaneously homooligomerize. widely used anti-cancer...
G-protein-coupled receptors (GPCRs) represent one of the largest gene families in human genome and have long been regarded as valuable targets for small-molecule drugs. The authors describe a new functional assay that directly monitors GPCR activation. It is based on interaction between beta-arrestin ligand-activated GPCRs uses enzyme fragment complementation technology. In this format, interest fused to small (approximately 4 kDa), optimized alpha peptide (termed ProLink) derived from...
Hetero-oligomeric complexes of G protein-coupled receptors (GPCRs) may represent novel therapeutic targets exhibiting different pharmacology and tissue- or cell-specific site action compared with receptor monomers homo-oligomers. An ideal tool for validating this concept pharmacologically would be a hetero-oligomer selective ligand. We set out to develop execute 1536-well high-throughput screen over 1 million compounds detect potential ligands using β-arrestin recruitment assay in U2OS cells...
G-protein-coupled receptor (GPCR) signaling is involved in a wide range of physiological processes and diseases, around one-half currently used drugs target GPCRs. Assays for the GPCRs have suffered from drawbacks, including low signal-to-noise, temporally transient signals, difficulty applying single assay to We developed set assays based on beta-galactosidase enzyme complementation live mammalian cells. previously described an GPCR activation by monitoring binding beta-arrestin receptor....
To determine the pharmacology of ETA- and ETB-mediated β-arrestin recruitment compare this to established human these receptors identify evidence for endothelin receptor biased signalling pathway specific blockade by antagonists. The ability ET-1, ET-2, ET-3, sarafotoxin 6b 6c activate ETA was determined in CHO-K1 cells. Affinities were obtained selective (BQ123, sitaxentan, ambrisentan), ETB (BQ788) mixed (bosentan) antagonists using ET-1 compared affinities competition experiments heart...
G-protein coupled receptors (GPCRs) are a versatile and ubiquitous family of membrane that transmit extracellular signals to mammalian cells constitute the most important class drug targets. Yet, sensitive specific methods lacking would allow quantitative comparisons pharmacologic properties these in physiological or pathological settings live animals. We sought overcome limitations by employing low affinity, reversible beta-galactosidase complementation quantify GPCR activation via...
G-protein-coupled receptors (GPCRs) have varying and diverse physiological roles, transmitting signals from a range of stimuli, including light, chemicals, peptides, mechanical forces. More than 130 GPCRs are orphan (i.e., their endogenous ligands unknown), representing large untapped reservoir potential therapeutic targets for pharmaceutical intervention in variety diseases. Current deorphanization approaches slow, laborious, usually require some in-depth knowledge about the receptor...
<h3>Introduction</h3> Baricitinib (BARI), an oral selective Janus kinase (JAK) 1/2 inhibitor, approved in the EU for moderate to severe active RA. <h3>Objectives</h3> To compare <i>in vitro</i> cellular pharmacology of BARI upadacitinib (ABT), filgotinib (FILGO), and tofacitinib (TOFA), three JAK inhibitors (JAKis) currently or clinical development. <h3>Methods</h3> Peripheral blood mononuclear cells from healthy donors (n=6–12) were incubated with different JAKis. After cytokine...
Background: everal JAKi are now used for the treatment of RA; approved doses include baricitinib (bari) 2- and/or 4-mg QD, tofacitinib (tofa) 5-mg BID, upadacitinib (upa) 15-mg QD. The JAK selectivity these agents is proposed to vary across class. Objectives: In vitro cellular pharmacology bari tofa, upa, and filgotinib (filgo) were compared. Methods: PBMCs from 6 healthy donors incubated with JAKis over a 7- 8-point concentration range. Following cytokine stimulation, levels pSTAT measured...
<h3>Background</h3> Baricitinib (bari) and tofacitinib (tofa) are selective JANUS kinase inhibitors (JAKi). In biochemical assays, bari exhibits greatest potency against JAK1 JAK2, whereas that for tofa is JAK3. <h3>Objectives</h3> To determine whether differences in JAKi specificity translate to cellular signal transduction leukocyte subpopulations stimulated by cytokines, indicated inhibition of transducers activator transcription (STATs) phosphorylation. <h3>Methods</h3> Peripheral blood...