A5032432916- Melanoma and MAPK Pathways
- Mass Spectrometry Techniques and Applications
- NF-κB Signaling Pathways
- Analytical Chemistry and Chromatography
- Metabolomics and Mass Spectrometry Studies
- Metal-Catalyzed Oxygenation Mechanisms
- Microbial bioremediation and biosurfactants
- Advanced Proteomics Techniques and Applications
- Biofuel production and bioconversion
- Computational Drug Discovery Methods
- Synthesis and biological activity
- Enzyme Structure and Function
- Microbial metabolism and enzyme function
- Radioactive element chemistry and processing
- CAR-T cell therapy research
- Biochemical and Molecular Research
- Research in Cotton Cultivation
- Glycosylation and Glycoproteins Research
- Lymphoma Diagnosis and Treatment
- Carbohydrate Chemistry and Synthesis
- Microbial Metabolic Engineering and Bioproduction
- Viral Infectious Diseases and Gene Expression in Insects
- Enzyme Production and Characterization
- Photosynthetic Processes and Mechanisms
- Atmospheric and Environmental Gas Dynamics
AbbVie (United States)
2016-2025
United States Military Academy
2012-2017
Merck & Co., Inc., Rahway, NJ, USA (United States)
2015-2016
University of Wisconsin–Madison
2007-2014
Great Lakes Bioenergy Research Center
2010-2011
Methicillin-resistant Staphylococcus aureus infection can be treated effectively by combining a β-lactam antibiotic with drug that targets FtsZ.
The cooperative assembly of FtsZ, the prokaryotic homologue tubulin, plays an essential role in cell division. FtsZ is a potential drug target, as illustrated by small-molecule cell-cycle inhibitor and antibacterial agent PC190723 that targets FtsZ. We demonstrate negatively modulates Staphylococcus aureus polymerization cooperativity reflected at lower concentrations without defined critical concentration. crystal structure S. FtsZ-PC190723 complex shows domain movement would stabilize...
Abstract RORγt is critical for the differentiation and proliferation of Th17 cells associated with several chronic autoimmune diseases. We report discovery a novel allosteric binding site on nuclear receptor RORγt. Co-crystallization ligand domain (LBD) series small-molecule antagonists demonstrates occupancy previously unreported pocket. Binding at this non-canonical induces an unprecedented conformational reorientation helix 12 in LBD, which blocks cofactor binding. The functional...
The adoption of mass spectrometry for high-throughput screening in drug discovery has become increasingly prevalent and enabled label-free against diverse targets. Cellular assays phenotypic screening, however, are primarily conducted by microscopy as there remain many challenges associated with conducting screens via ultra-high throughput spectrometry. Following a simple on-plate extraction, infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) was employed to...
A microarray study of chemostat growth on insoluble cellulose or soluble cellobiose has provided substantial new information Clostridium thermocellum gene expression. This is the first comprehensive examination expression in C. under defined conditions. Expression was detected from 2,846 3,189 genes, and regression analysis revealed 348 genes whose changes patterns were rate and/or substrate dependent. Successfully modeled included those for scaffoldin cellulosomal enzymes, intracellular...
The MAPK pathway is frequently activated in many human cancers, particularly melanomas. A single-nucleotide mutation BRAF resulting the substitution of glutamic acid for valine (V(600E)) causes constitutive activation downstream pathway. Selective and MEK inhibitor therapies have demonstrated remarkable antitumor responses BRAF(V600) (E)-mutant melanoma patients. However, initial tumor shrinkage transient vast majority patients develop resistance. We previously reported that SCH772984, an...
Infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) mass spectrometry is an ambient-direct sampling method that being developed for high-throughput, label-free, biochemical screening of large-scale compound libraries. Here, we report the development ultra-high-throughput continuous motion IR-MALDESI approach capable acquiring data at rates up to 22.7 samples per second in a 384-well microtiter plate. At top speed, less than 1% analyte carryover observed from...
Infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) is a hybrid, ambient source that combines the advantages of and desorption/ionization, making it versatile tool for both high-throughput screening (HTS) mass spectrometry imaging (MSI) studies. To expand capabilities IR-MALDESI source, an entirely new architecture was designed to overcome key limitations previous source. This next-generation (NextGen) features vertically mounted IR-laser, planar translation stage...
Mass spectrometry (MS) is the primary analytical tool used to characterize proteins within biopharmaceutical industry. Electrospray ionization (ESI) coupled liquid chromatography (LC) current gold standard for intact protein analysis. However, inherent speed limitations of LC/MS prevent analysis large sample numbers (>1000) in a day. Infrared matrix-assisted laser desorption electrospray (IR-MALDESI-MS), an ambient MS technology, has recently been established as platform high-throughput...
Infrared matrix-assisted laser desorption ionization (IR-MALDESI) is a hybrid mass spectrometry source that combines the benefits of electrospray (ESI) and (MALDI) making it great analytical tool for high-throughput screening (HTS) analyses. IR-MALDESI coupled to an Orbitrap Exploris 240 spectrometer utilizes bent quadrupole (C-trap) inject accumulated ions into high-field analyzer. Here, we present study on optimized C-trap timing HTS analyses by spectrometry. The between initial ion...
Mass spectrometry (MS) detection can offer unmatched selectivity and sensitivity. The use of MS without chromatography greatly increases the throughput, making it suitable for high throughput screening. However, trade-offs direct need to be carefully evaluated along with development novel strategies ensure successful implementation. In this review, we will discuss pros cons chromatography-free some currently used future technologies being investigated enable high-throughput MS.
Mass spectrometry (MS) can provide high sensitivity and specificity for biochemical assays without the requirement of labels, eliminating risk assay interference. However, its use had been limited to lower-throughput due need chromatography overcome ion suppression from sample matrix. Direct analysis has potential throughput if is sufficient despite presence a Here, we report demonstrate novel direct high-throughput MS system based on infrared matrix-assisted desorption electrospray...
Cellular pharmacodynamic assays are crucial aspects of lead optimization programs in drug discovery. These sometimes difficult to develop, oftentimes distal from the target and frequently low throughput, which necessitates their incorporation discovery funnel later than desired. The earlier direct modulation a can be established, fewer resources wasted on compounds that acting via an off-target mechanism. Mass spectrometry is versatile tool often used for direct, proximal cellular assay...
Abstract The activated B cell (ABC) subset of diffuse large B-cell lymphoma (DLBCL) is characterized by chronic receptor signaling and associated with poor outcomes when treated standard therapy. In ABC-DLBCL, MALT1 a core enzyme that constitutively stimulation the or gain-of-function mutations in upstream components pathway, making it an attractive therapeutic target. We discovered novel small-molecule inhibitor, ABBV-MALT1, potently shuts down selectively ABC-DLBCL preclinical models...
The primary objective of early drug discovery is to associate druggable target space with a desired phenotype. inability efficiently these often leads failure in the process. In this proof-of-concept study, most tractable starting points for within NF-κB pathway model system were identified by integrating affinity selection-mass spectrometry (AS-MS) functional cellular assays. AS-MS platform Automated Ligand Identification System (ALIS) was used rapidly screen 15 proteins parallel against...
Abstract Productive biomolecular recognition requires exquisite control of affinity and specificity. Accordingly, nature has devised many strategies to achieve proper binding interactions. Bacterial multicomponent monooxygenases provide a fascinating example, where diiron hydroxylase must reversibly interact with both ferredoxin catalytic effector in order electron transfer O 2 activation during catalysis. Because these two accessory proteins have distinct structures, because the...
Deconvolution from intact protein mass-to-charge spectra to mass is essential generate interpretable data for spectrometry (MS) platforms coupled ionization sources that produce multiply charged species. Infrared matrix-assisted laser desorption electrospray (IR-MALDESI) can be used analyze proteins in multiwell microtiter plates with speed matching small molecule analyses (at least 1 Hz). However, the lack of compatible deconvolution software has limited its use high-throughput screening...
Clostridium thermocellum is a cellulosome-producing bacterium that able to efficiently degrade and utilize cellulose as sole carbon source. Cellobiose phosphorylase (CBP) plays critical role in degradation by catalyzing the reversible phosphate-dependent hydrolysis of cellobiose, major product degradation, into α-D-glucose 1-phosphate D-glucose. CBP from C. modular enzyme composed four domains [N-terminal domain, helical linker, (α/α)(6)-barrel domain C-terminal domain] member glycoside...
The active site residue Thr-201 in toluene 4-monooxygenase hydroxylase (T4moH) has a structural counterpart the sites of all diiron monooxygenases. Thus, our previous finding that mutation this to Ala, Gly, or Ser had no impact on steady-state catalysis coupling was surprising. In work, we provide kinetic, biochemical, and evidence one role may be stabilize peroxo-level intermediate during enzyme catalysis. During reactions absence substrate, T201 T4moH slowly consumed O(2) but only...
Crystal structures of toluene 4-monooxygenase hydroxylase in complex with reaction products and effector protein reveal active site interactions leading to regiospecificity. Complexes phenolic yield an asymmetric μ-phenoxo-bridged diiron center a shift ligand E231 into hydrogen bonding position conserved T201. In contrast, complexes inhibitors p-NH2-benzoate p-Br-benzoate showed μ-1,1 coordination carboxylate oxygen between the iron atoms only partial E231. Among residues, F176 trapped...
High-throughput screening (HTS) is a critical step in the drug discovery process. However, most mass spectrometry (MS)-based HTS methods require sample cleanup steps prior to analysis. In this work we present utility of infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) for monitoring an enzymatic reaction directly from biological buffer system with no and at high throughput.
Introduction Ultra-high-throughput mass spectrometry, uHT-MS, is a technology that utilizes ionization and sample delivery technologies optimized to enable sampling from well plates at > 1 per second. These do not need chromatographic separation step can be utilized in wide variety of assays detect broad range analytes including small molecules, lipids, proteins.
Toluene 4-monooxygenase catalyzes the NADH- and O2-dependent hydroxylation of toluene to form p-cresol. The four-protein complex consists a diiron hydroxylase, an oxidoreductase, catalytic effector protein, Rieske-type ferredoxin (T4moC). Phylogenetic analysis suggests that T4moC is part clade specialized for reaction with hydroxylases, possibly reflected in conservation W69, whose indole side chain makes close contacts bridging sulfide. In order further investigate possible origins this...