A5040662926- Protein Degradation and Inhibitors
- Monoclonal and Polyclonal Antibodies Research
- Multiple Myeloma Research and Treatments
- Ubiquitin and proteasome pathways
- Advanced Biosensing Techniques and Applications
- Protein purification and stability
- Peptidase Inhibition and Analysis
- Advanced Proteomics Techniques and Applications
- Biotin and Related Studies
- Molecular Biology Techniques and Applications
- Click Chemistry and Applications
- Histone Deacetylase Inhibitors Research
- RNA modifications and cancer
- Bacterial Genetics and Biotechnology
- HIV/AIDS drug development and treatment
- Viral Infectious Diseases and Gene Expression in Insects
- Mass Spectrometry Techniques and Applications
- CAR-T cell therapy research
- HER2/EGFR in Cancer Research
- DNA Repair Mechanisms
- Genomics and Chromatin Dynamics
- RNA and protein synthesis mechanisms
- Advanced Fluorescence Microscopy Techniques
- Glycosylation and Glycoproteins Research
- Biosimilars and Bioanalytical Methods
Promega (United States)
2016-2025
LI-COR Biosciences (United States)
2002
University of Wisconsin–Madison
1994-1998
We have designed a modular protein tagging system that allows different functionalities to be linked onto single genetic fusion, either in solution, living cells, or chemically fixed cells. The tag (HaloTag) is modified haloalkane dehalogenase covalently bind synthetic ligands (HaloTag ligands). comprise chloroalkane linker attached variety of useful molecules, such as fluorescent dyes, affinity handles, solid surfaces. Covalent bond formation between the and highly specific, occurs rapidly...
Bromodomain inhibitors revisited and extraterminal domain (BET) proteins contribute to the pathogenesis of cancer immune diseases through their effects on transcriptional regulation. BET contain two nearly identical bromodomains, BD1 BD2, structural modules that have attracted great interest as targets for drug development. First-generation drugs inhibited both BD2 showed promising therapeutic activity in preclinical models but proved be less efficacious clinical trials. Gilan et al. took a...
A new generation of heterobifunctional small molecules, termed proteolysis targeting chimeras (PROTACs), targets proteins for degradation through recruitment to E3 ligases and holds significant therapeutic potential. Despite numerous successful examples, PROTAC molecule development remains laborious unpredictable, involving testing compounds end-point activity at fixed times concentrations without resolving or optimizing the important biological steps required process. Given complexity...
Abstract The histone acetyltransferases CBP/p300 are involved in recurrent leukemia-associated chromosomal translocations and key regulators of cell growth. Therefore, efforts to generate inhibitors clinical value. We developed a specific potent acetyl-lysine competitive protein–protein interaction inhibitor, I-CBP112, that targets the bromodomains. Exposure human mouse leukemic lines I-CBP112 resulted substantially impaired colony formation induced cellular differentiation without...
PROteolysis TArgeting Chimeras (PROTACs) are hetero-bifunctional small molecules that can simultaneously recruit target proteins and E3 ligases to form a ternary complex, promoting protein ubiquitination degradation via the Ubiquitin-Proteasome System (UPS). PROTACs have gained increasing attention in recent years due certain advantages over traditional therapeutic modalities enabling targeting of previously "undruggable" proteins. To better understand mechanism PROTAC-induced Target Protein...
Molecular-glue degraders are small molecules that induce a specific interaction between an E3 ligase and target protein, resulting in the proteolysis. The discovery of molecular glue currently relies mostly on screening approaches. Here, we describe library cereblon (CRBN) ligands against panel patient-derived cancer cell lines, leading to SJ7095, potent degrader CK1α, IKZF1 IKZF3 proteins. Through structure-informed exploration structure activity relationship (SAR) around this molecule...
In April 2016, the Food and Drug Administration approved first biosimilar monoclonal antibody (mAb), Inflectra/Remsima (Celltrion), based off original product Remicade (infliximab, Janssen). Biosimilars promise significant cost savings for patients, but unavoidable differences between innovator copycat biologics raise questions regarding interchangeability. this study, Remsima were examined by native mass spectrometry, ion mobility, quantitative peptide mapping. The levels of oxidation,...
Identification of proteins resolved by SDS-PAGE depends on robust in-gel protein digestion and efficient peptide extraction, requirements that are often difficult to achieve. A lengthy laborious procedure is an additional challenge identification in gel. We show here with the use mass spectrometry compatible surfactant sodium 3-((1-(furan-2-yl)undecyloxy)carbonylamino)propane-1-sulfonate, challenges effectively addressed. Peptide quantitation based stable isotope labeling showed induced...
PROTACs usually occupy physicochemical space outside the one defined by classical drug-like molecules, which often presents considerable challenges in their optimization and development for oral administration. We have previously reported phenyl glutarimide (PG)-based BET PROTAC SJ995973, with improved overall vitro degradation antiproliferative activities compared to its direct thalidomide-based analogue dBET1, but similarly poor vivo pharmacokinetic profile. To further demonstrate PG...
Understanding protein function and interaction is central to the elucidation of biological processes. Systematic analysis interactions have shown that eukaryotic proteome highly interconnected frequently depends on orchestrated action many proteins. Perturbation these functions or can lead various disease states pharmacologic intervention result in corrective therapies. The fact proteins rarely act isolation, but rather comprise complex machines stably and/or transiently interact with...
Receptor-mediated antibody internalization is a key mechanism underlying several anti-cancer therapeutics. Delivering highly toxic drugs to cancer cells, as in the case of drug conjugates (ADCs), efficient removal surface receptors from cells and changing pharmacokinetics profile are some ways that impacts therapeutic efficacy antibodies. Over years, techniques have been used study including radiolabels, fluorescent microscopy, flow cytometry cellular toxicity assays. While these methods...
Characterization of asparagine deamidation and aspartic acid isomerization is an important aspect biotherapeutic protein analysis due to the potential negative effect these modifications on drug efficacy stability. Succinimide has long been known be intermediate product isomerization, but despite key role succinimide in reactions, its remains challenging instability. We have developed a paradigm which two interlinked analytical methods are used develop optimized approach analyze succinimide....
Targeted protein degradation using heterobifunctional proteolysis-targeting chimera (PROTAC) compounds, which recruit E3 ligase machinery to a target protein, is increasingly becoming an attractive pharmacologic strategy. PROTAC compounds are often developed from existing inhibitors, and assessing selectivity critical for understanding on-target off-target degradation. We present here in-depth kinetic study of the pan-kinase PROTAC, TL12-186, applied 16 members cyclin-dependent kinase (CDK)...
Abstract To assess the role of a protein, protein loss phenotypic studies can be used, most commonly through mutagenesis RNAi or CRISPR knockout. Such have been critical for understanding function and identification putative therapeutic targets numerous human disease states. However, these methodological approaches present challenges because they are not easily reversible, if an essential gene is targeted, associated cell viability potentially hinder further studies. Here we reversible...
Proteolysis targeting chimera (PROTAC) degraders are typically bifunctional with one moiety of an E3 ligase ligand connected to target protein via a linker. While augmented valency has been shown trivalent PROTACs two binding sites within given protein, or used recruit different targets, the possibility recruiting ligases same compound not demonstrated. Here we present dual-ligase recruitment as strategy enhance targeted degradation. We designed heterotrivalent composed CRBN, VHL and BET...