Publications by authors named "Jordan M Mattheisen"
Article Synopsis
- Methods in chemical biology, specifically genetic code expansion (GCE), have greatly improved the study of integral membrane proteins by allowing the incorporation of noncanonical amino acids (ncAAs).
- GCE is particularly challenging for membrane proteins due to their unique characteristics and low expression levels, requiring effective use of mammalian cell cultures for functional expression.
- Recent advancements include engineered AARS/tRNA pairs for better performance in mammalian cells, bioorthogonal reactions for attaching probes to live cell membrane proteins, and an expanded selection of ncAAs for in-depth analysis of protein structure and dynamics.
Objectives: To develop a biological diary (CoronaCal) that allows anyone in the community to collect and store serial saliva samples and chart symptoms on ordinary printer paper.
Methods: Diaries were analyzed for the presence of SARS-CoV-2 RNA using established polymerase chain reaction (PCR) procedures. CoronaCal diaries were distributed to volunteer subjects in the community during the peak of the COVID-19 outbreak in New York.
We report a bioluminescence resonance energy transfer (BRET) assay to quantitate the fraction of an engineered membrane protein at the cell surface versus inside the cell. As test cases, we engineered two different G protein-coupled receptors (GPCRs) in which a NanoLuc luciferase (NLuc) and a HaloTag are fused to the extracellular amino-terminal tail of the receptors. We then employed a pulse-chase labeling approach relying on two different fluorescent dyes with distinctive cell permeability properties.
View Article and Find Full Text PDFG protein-coupled receptors (GPCRs) modulate diverse cellular signaling pathways and are important drug targets. Despite the availability of high-resolution structures, the discovery of allosteric modulators remains challenging due to the dynamic nature of GPCRs in native membranes. We developed a strategy to covalently tether drug fragments adjacent to allosteric sites in GPCRs to enhance their potency and enable fragment-based drug screening in cell-based systems.
View Article and Find Full Text PDFFor use in site-specific bioorthogonal labeling of expressed G protein-coupled receptors (GPCRs) in live cells, we developed a luciferase-based reporter assay. The assay was used to compare amber codon suppression efficiency, receptor functionality, and efficiency of different bioorthogonal labeling chemistries. We used the assay system to compare side-by-side the efficiency of incorporation of three different noncanonical amino acids [4-azido-l-phenylalanine (azF), cyclopropene-l-lysine (CpK), and trans-cyclooct-2-en-l-lysine (TCOK)] at three different sites on a GPCR using three different genetic code expansion plasmid systems.
View Article and Find Full Text PDFUveal melanoma is the most common eye cancer in adults and is clinically and genetically distinct from skin cutaneous melanoma. In a subset of cases, the oncogenic driver is an activating mutation in CYSLTR2, the gene encoding the G protein-coupled receptor cysteinyl-leukotriene receptor 2 (CysLTR2). The mutant CYSLTR2 encodes for the CysLTR2-L129Q receptor, with the substitution of Leu to Gln at position 129 (3.
View Article and Find Full Text PDFArticle Synopsis
- Researchers developed a new method to create biosensors in E. coli that can detect environmental chemicals by stabilizing a transcriptional activator in response to specific ligands.
- They constructed a biosensor by fusing the lac repressor protein (LacI) to a DNA-binding domain and a transcription-activating domain, achieving a significant 7-fold increase in bacterial growth rate in response to the ligand IPTG.
- The strategy, which involves mutating the lacI gene and testing its efficacy, was also successfully applied to engineer another protein, MphR, demonstrating its potential for wider use in creating biosensors from natural proteins.