A5083510496- Melanoma and MAPK Pathways
- Protein Kinase Regulation and GTPase Signaling
- Computational Drug Discovery Methods
- Synthesis and biological activity
- Receptor Mechanisms and Signaling
- Advanced Breast Cancer Therapies
- Ferroptosis and cancer prognosis
- Histone Deacetylase Inhibitors Research
- Cancer Mechanisms and Therapy
- SARS-CoV-2 and COVID-19 Research
- Cancer-related Molecular Pathways
- Cytokine Signaling Pathways and Interactions
- Cancer, Hypoxia, and Metabolism
- Magnesium in Health and Disease
- Inflammatory mediators and NSAID effects
- X-ray Diffraction in Crystallography
- Crystallization and Solubility Studies
- Synthesis and Biological Evaluation
- Ubiquitin and proteasome pathways
- Chemical Synthesis and Analysis
- Click Chemistry and Applications
- Advanced Biosensing Techniques and Applications
- Mass Spectrometry Techniques and Applications
- CAR-T cell therapy research
- Monoclonal and Polyclonal Antibodies Research
The University of Texas at Austin
2016-2025
Minia University
2013-2019
Austin College
2014-2017
King Abdulaziz University
2014
Institute of Medicinal Plant Development
2011
To address the ongoing SARS-CoV-2 pandemic and prepare for future coronavirus outbreaks, understanding protective potential of epitopes conserved across variants lineages is essential. We describe a highly conserved, conformational S2 domain epitope present only in prefusion core β-coronaviruses: apex residues 980–1006 flexible hinge. Antibody RAY53 binds native hinge MERS-CoV spikes on surface mammalian cells mediates antibody-dependent cellular phagocytosis cytotoxicity against spike...
Eukaryotic elongation factor 2 kinase (eEF-2K), through its phosphorylation of (eEF2), provides a mechanism by which cells can control the rate phase protein synthesis. The activity eEF-2K is increased in rapidly proliferating malignant cells, inhibited during mitosis, and may contribute to promotion autophagy response anti-cancer therapies. purpose this study was examine therapeutic potential targeting breast cancer tumors. Through systemic administration liposomal siRNA (twice week, i.v....
Mitogen-activated protein (MAP) kinases are responsible for many cellular functions, and their malfunction manifests itself in several human diseases. Usually, monitoring the phosphorylation states of MAP vitro requires preparation purification proteins or Western blotting. Herein, we report an array sensing approach differentiation phosphorylated counterparts vitro. This technique utilizes a library differential receptors created situ containing peptides known affinity to kinases,...
The SARS-CoV-2 spike protein is a critical component of vaccines and target for neutralizing monoclonal antibodies (nAbs). Spike also undergoing immunogenic selection with variants that increase infectivity partially escape convalescent plasma. Here, we describe Display, high-throughput platform to rapidly characterize glycosylated ectodomains across multiple coronavirus-family proteins. We assayed ∼200 variant spikes their expression, ACE2 binding, recognition by 13 nAbs. An alanine scan...
193-nm ultraviolet photodissociation (UVPD) was implemented to sequence singly and multiply charged peptide anions. Upon dissociation by this method, a-/x-type, followed d w side-chain loss ions, were the most prolific abundant often yielding 100% coverage. The behavior of anions significantly different with higher precursors more ions; however, all charge states investigated (1- through 3-) produced rich diagnostic information. UVPD at 193 nm also shown successfully differentiate pinpoint...
Arrestins make up a small family of proteins with four mammalian members that play key roles in the regulation multiple G protein-coupled receptor-dependent and -independent signaling pathways. Although arrestins were reported to serve as scaffolds for MAP kinase cascades, promoting activation JNK3, ERK1/2, p38, molecular mechanisms involved not elucidated, even direct binding kinases was never demonstrated. Here, using purified proteins, we show both nonvisual directly bind JNK3α2 its...
Eukaryotic elongation factor 2 kinase (eEF-2K) is an atypical protein regulated by Ca2+ and calmodulin (CaM). Its only known substrate eukaryotic (eEF-2), whose phosphorylation eEF-2K impedes global synthesis. To date, the mechanism of autophosphorylation has not been fully elucidated. investigate autophosphorylation, human was coexpressed with λ-phosphatase purified from bacteria in a three-step protocol using CaM affinity column. Purified induced to autophosphorylate incubation Ca2+/CaM...
Three-kinase mitogen-activated protein kinase (MAPK) signaling cascades are present in virtually all eukaryotic cells. MAPK organized by scaffold proteins, which assemble cognate kinases into productive complexes. Arrestin-3 facilitates JNK activation cells, and a short 25-residue arrestin-3 peptide was identified as the critical JNK3-binding element. Here we demonstrate that this also binds MKK4, MKK7, ASK1, upstream JNK3-activating kinases. This is sufficient to enhance JNK3 activity A...
ABSTRACT Three pathogenic human coronaviruses have emerged within the last 20 years, with SARS-CoV-2 causing a global pandemic. Although therapeutic antibodies targeting spike currently focus on poorly conserved receptor-binding domain, essential neutralizing epitopes more S2 domain may provide broader protection. We report an antibody binding epitope in pre-fusion core of MERS-CoV, SARS-CoV and domains. Antibody 3A3 binds conformational ~2.5 nM affinity neutralizes from SARS-CoV, variants...
Evidence that elongation factor 2 kinase (eEF-2K) has potential as a target for anticancer therapy and possibly the treatment of depression is emerging. Here steady-state kinetic mechanism eEF-2K presented using peptide substrate shown to conform an ordered sequential with ATP binding first. Substrate inhibition by was observed revealed be competitive ATP, explaining mechanism. Several small molecules are reported inhibit activity most notable being histidine inhibitor NH125, which been used...
Abstract Collagen lysyl hydroxylases (LH1-3) are Fe 2+ - and 2-oxoglutarate (2-OG)-dependent oxygenases that maintain extracellular matrix homeostasis. High LH2 levels cause stable collagen cross-link accumulations promote fibrosis cancer progression. However, developing LH antagonists will require structural insights. Here, we report a 2 Å crystal structure X-ray scattering on dimer assemblies for the domain of L230 in Acanthamoeba polyphaga mimivirus . Loop residues double-stranded β-helix...
Significance Scaffold proteins are key regulators of signal transduction that bind multiple a signaling pathway. This simple tethering model explains how the scaffold colocalizes components for propagation, but it is unclear whether scaffolds ensure amplification. Here, we use arrestin-3 and ASK1-MKK4/7-JNK3 kinase cascade as system to explore scaffolding amplifies signaling. Our studies suggest scaffold-dependent amplification may be conceptually applicable other pathways.
Protein kinases are therapeutic targets for many human diseases, but the lack of user-friendly quantitative assays limits ability to follow activities numerous at once (multiplexing). To develop such an assay, we report array sulfonamido-oxine (SOX)-labeled peptides showing cross-reactivity different mitogen-activated protein (MAPKs) use in a differential sensing scheme. We first verified using linear discriminant analysis that could differentiate MAPK isoforms. Then, principal component...
Despite their lack of selectivity toward c-Jun N-terminal kinase (JNK) isoforms, peptides derived from the JIP (JNK Interacting Protein) scaffolds linked to cell-penetrating peptide TAT are widely used investigate JNK-mediated signaling events. To engineer an isoform-selective inhibitor, several JIP-based sequences were designed and tested. A sequence connected through a flexible linker either N-terminus inverted (JIP(10)-Δ-TAT(i)) or poly arginine (JIP(10)-Δ-R(9)) enabled potent inhibition...
The mitogen-activated protein (MAP) kinase ERK2 contains recruitment sites that engage canonical and noncanonical motifs found in a variety of upstream kinases, regulating phosphatases downstream targets. Interactions involving two these sites, the D-recruitment site (DRS) F-recruitment (FRS), have been shown to play key role signal transduction by ERK/MAP kinases. dynamic nature events makes NMR uniquely suited provide significant insight into interactions. While studies kinases general...
Abstract Five SOX peptides are used to classify the MAPK groups and isoforms thereof using chemometrics. The score plots show excellent classification accuracy, while support vector machine analysis leads quantification of ERK an inhibitor concentration in kinase mixtures. Examination loading reveals cross‐reactivity among peptides, some unexpected surprises.
// Nancy D. Ebelt 1, 2 , Tamer S. Kaoud 2, 4 Ramakrishna Edupuganti Sabrina Van Ravenstein Kevin N. Dalby and Carla L. Den Berg 3 1 Institute of Cellular & Molecular Biology, University Texas at Austin, Dell Pediatric Research Institute, TX 78723, USA Division Chemical Biology Medicinal Chemistry, College Pharmacy, 78712, Pharmacology Toxicology, Department Faculty Minia University, El-Minia 61519, Egypt Correspondence to: Berg, email: carla.vandenberg@austin.utexas.edu Keywords: triple...