A5010523573- Monoclonal and Polyclonal Antibodies Research
- CAR-T cell therapy research
- Biosimilars and Bioanalytical Methods
- Cancer Mechanisms and Therapy
- Wnt/β-catenin signaling in development and cancer
- Cancer Cells and Metastasis
- Bacterial Genetics and Biotechnology
- Peptidase Inhibition and Analysis
- Cancer-related gene regulation
- HER2/EGFR in Cancer Research
- Microtubule and mitosis dynamics
- Chemical Synthesis and Analysis
- Cytomegalovirus and herpesvirus research
- Protein Kinase Regulation and GTPase Signaling
- Multiple Myeloma Research and Treatments
- Chemical Reactions and Isotopes
- RNA and protein synthesis mechanisms
- Drug Transport and Resistance Mechanisms
- Trace Elements in Health
- Lipid Membrane Structure and Behavior
- Protein Hydrolysis and Bioactive Peptides
- Enzyme Structure and Function
- Toxin Mechanisms and Immunotoxins
- Protein Degradation and Inhibitors
- CRISPR and Genetic Engineering
Novartis (United States)
2012-2025
Novartis (China)
2023
Boston Biomedical Research Institute
2023
Novartis (Switzerland)
2012-2013
Pfizer (United States)
2007-2011
Duke University Hospital
1997-2002
Duke Medical Center
1997-2002
The Ohio State University
1992-2000
Duke University
1997
We report that a thiol leader peptidase, produced by replacing the critical serine at position 90 with cysteine residue, is enzymatically active. In contrast to wild-type enzyme can be inactivated N-ethylmaleimide, cysteine-specific reagent. This strongly suggests involved in catalysis and located active site. Of three conserved basic residues signal peptidase family, only lysine 145 appears for catalysis; when was mutated an alanine K145A protein inactive both vitro vivo. A control...
Multiple myeloma has a continued need for more effective and durable therapies. B cell maturation antigen (BCMA), plasma surface member of the tumor necrosis factor (TNF) receptor superfamily, is an attractive target immunotherapy multiple due to its high prevalence on malignant cells. The current work details pre-clinical evaluation BCMA expression development chimeric (CAR) targeting this using fully human single chain variable fragment (scFv). We demonstrate that prevalently, but variably...
Lipoprotein lipase (LPL) plays a central role in triglyceride (TG) metabolism. By catalyzing the hydrolysis of TGs present TG-rich lipoproteins (TRLs), LPL facilitates TG utilization and regulates circulating TRL concentrations. Until very recently, structural information for was limited to homology models, presumably due propensity unfold aggregate. coexpressing with soluble variant its accessory protein glycosylphosphatidylinositol-anchored high-density lipoprotein binding 1 (GPIHBP1)...
P-cadherin (pCAD) and LI-cadherin (CDH17) are cell-surface proteins belonging to the cadherin superfamily that both highly expressed in colorectal cancer. This co-expression profile presents a novel attractive opportunity for dual targeting approach using an antibody–drug conjugate (ADC). In this study, we used unique avidity-driven vitro screening generate pCAD x CDH17 bispecific antibodies selectively target cells expressing antigens over only or CDH17. Based on binding inhibition of cell...
CRISPR/Cas9 mediated gene editing of patient-derived hematopoietic stem and progenitor cells (HSPCs) ex vivo followed by autologous transplantation the edited HSPCs back to patient can provide a potential cure for monogenic blood disorders such as β-hemoglobinopathies. One challenge this strategy is efficient delivery ribonucleoprotein (RNP) complex, consisting purified Cas9 protein guide RNA, into HSPCs. Because β-hemoglobinopathies are most prevalent in developing countries, it desirable...
Protein farnesyltransferase (FTase) catalyzes the modification by a farnesyl lipid of Ras and several other key proteins involved in cellular regulation. Previous studies on this important enzyme have indicated that product dissociation is rate-limiting step catalysis. A detailed examination has now been performed, results provide surprising insights into mechanism enzyme. Examination binding farnesylated peptide to free revealed affinity ∼1 μM. However, analysis release under single...
Despite an improving therapeutic landscape, significant challenges remain in treating the majority of patients with advanced ovarian or renal cancer. We identified cell-cell adhesion molecule cadherin-6 (CDH6) as a lineage gene having differential expression and kidney cancers. HKT288 is optimized CDH6-targeting DM4-based antibody-drug conjugate (ADC) developed for treatment these diseases. Our study provides mechanistic evidence supporting importance linker choice optimal antitumor activity...
Escherichia coli leader peptidase, which catalyzes the cleavage of signal peptides from pre-proteins, is an essential, integral membrane serine peptidase that has its active site residing in periplasmic space. It contains a conserved lysine residue been proposed to act as general base, abstracting proton side chain hydroxyl group nucleophilic 90. To help elucidate role essential 145 activity E. we have combined site-directed mutagenesis and chemical modification methods introduce unnatural...
JNK1 (c-Jun N-terminal kinase 1) plays a crucial role in the regulation of obesity-induced insulin resistance and is implicated pathology Type 2 diabetes. Its partner, JIP1 (JNK-interacting protein 1), serves scaffolding function that facilitates activation by MKK4 [MAPK (mitogen-activated kinase) 4] MKK7 (MAPK 7). For example, reduced JNK are observed JIP1-deficient mice. On basis vivo efficacy cell-permeable JIP peptide, JIP-JNK interaction appears to be potential target for inhibition....
ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTCharacterization of a soluble, catalytically active form Escherichia coli leader peptidase: requirement detergent or phospholipid for optimal activityWilliam Tschantz, Mark Paetzel, Guoqing Cao, Dominic Suciu, Masayori Inouye, and Ross E. DalbeyCite this: Biochemistry 1995, 34, 12, 3935–3941Publication Date (Print):March 28, 1995Publication History Published online1 May 2002Published inissue 28 March 1995https://doi.org/10.1021/bi00012a010RIGHTS &...
Antibody–drug conjugates (ADCs) are an established modality that allow for targeted delivery of a potent molecule, or payload, to desired site action. ADCs, wherein the payload is protein degrader, emerging area in field. Herein we describe our efforts delivering Bruton's tyrosine kinase (BTK) bifunctional degrader 1 via CD79b mAb (monoclonal antibody) where linked at ligase binding portion cleavable linker mAb. The resulting 3 and 4, exhibit vitro degradation cytotoxicity comparable with 1,...
The removal of endotoxin from protein solutions and its prevention are key to the success recombinant production due possible pyogenic response in mammals caused by contaminated samples. In pre-clinical situation, is often carried out a non-good manufacturing practice (GMP) setting, utilizing bacterial DNA for transient transfection non-validated cleaning techniques. Here, we present our findings evaluating various options removal, propose strategies with emphasis on chromatographic...
The cell surface glycoprotein P-cadherin is highly expressed in a number of malignancies, including those arising the epithelium bladder, breast, esophagus, lung, and upper aerodigestive system. PCA062 specific antibody-drug conjugate that utilizes clinically validated SMCC-DM1 linker payload to mediate potent cytotoxicity lines expressing high levels vitro, while displaying no activity P-cadherin-negative lines. High necessary, but not sufficient, cytotoxicity. In vivo, demonstrated serum...
Prenylated proteins contain either a 15-carbon farnesyl or 20-carbon geranylgeranyl isoprenoid covalently attached via thioether bond to cysteine residue at near their C terminus. As prenylated comprise up 2% of the total protein in eukaryotic cells, and is stable modification, degradation raises metabolic challenge cells. A lysosomal enzyme termed prenylcysteine lyase has been identified that cleaves prenylcysteines an unidentified product. Here we show product C-1 aldehyde moiety (farnesal...
Type I signal peptidases are integral membrane proteins that function to remove peptides from secreted and proteins. These enzymes carry out catalysis using a serine/lysine dyad instead of the prototypical serine/histidine/aspartic acid triad found in most serine proteases. Site-directed scanning mutagenesis was used obtain qualitative assessment which residues fifth conserved region, Box E, <i>Escherichia coli</i> peptidase critical for maintaining functional enzyme. First, we find there is...
CD3-bispecific antibodies represent an important therapeutic strategy in oncology. These molecules work by redirecting cytotoxic T cells to antigen-bearing tumor cells. Although have been developed for several clinical indications, cases of cancer-derived resistance are emerging limitation the more generalized application these molecules. Here, we devised whole-genome CRISPR screens identify cancer mechanisms across multiple targets and types. By validating screen hits, found that deficiency...
Prenylated proteins contain either a 15-carbon farnesyl or 20-carbon geranylgeranyl isoprenoid covalently attached to cysteine residues at near their C terminus. These constitute up 2% of total cellular protein in eukaryotic cells. The degradation prenylated raises metabolic challenge the cell, because thioether bond modified is quite stable. We recently identified and isolated an enzyme termed prenylcysteine lyase that cleaves free product (Zhang, L., Tschantz, W. R., Casey, P. J. (1997)...
Abstract Leader peptidase, a novel serine protease in Escherichia coli , catalyzes the cleavage of amino‐terminal leader sequences from exported proteins. It is an integral membrane protein containing two transmembrane segments with its carboxy‐terminal catalytic domain residing periplasmic space. Here, we report procedure for purification and crystallization soluble non‐membrane‐bound form peptidase (Δ2‐75). Crystals were obtained by sitting‐drop vapor diffusion technique using ammonium...