A5018018134- Chemical Synthesis and Analysis
- Click Chemistry and Applications
- Monoclonal and Polyclonal Antibodies Research
- RNA and protein synthesis mechanisms
- Biochemical and Structural Characterization
- Advanced biosensing and bioanalysis techniques
- Antimicrobial Resistance in Staphylococcus
- HIV Research and Treatment
- Prion Diseases and Protein Misfolding
- Innovative Microfluidic and Catalytic Techniques Innovation
- Peptidase Inhibition and Analysis
- Synthesis and Biological Evaluation
- RNA Research and Splicing
- Signaling Pathways in Disease
- Genetics, Bioinformatics, and Biomedical Research
- Glycosylation and Glycoproteins Research
- Protein Degradation and Inhibitors
- RNA modifications and cancer
- RNA Interference and Gene Delivery
Harvard University
2015-2022
Broad Institute
2017-2020
Howard Hughes Medical Institute
2015-2018
Site-specific chemical modification of proteins can assist in the study their function. Furthermore, these methods are essential to develop biologicals for diagnostic and therapeutic use. Standard protein engineering protocols recombinant expression enable production with short peptide tags recognized by enzymes capable site-specific modification. We report here application two orthogonal specificity, sortase A butelase 1, prepare non-natural C-to-C fusion proteins. Using enzymes, we further...
We previously reported interaction determination using unpurified proteins (IDUP), a method to selectively amplify DNA sequences encoding ligand:target pairs from mixture of DNA-linked small molecules and protein targets in cell lysates. In this study, we applied IDUP libraries DNA-encoded bioactive compounds DNA-tagged human kinases identify ligand:protein binding partners out 32 096 possible combinations single solution-phase library × experiment. The results recapitulated known...
Abstract Although cyclophilins are attractive targets for probing biology and therapeutic intervention, no subtype-selective cyclophilin inhibitors have been described. We discovered novel from the in vitro selection of a DNA-templated library 256,000 drug-like macrocycles D (CypD) affinity. Iterated macrocycle engineering guided by ten X-ray co-crystal structures yielded potent selective (half maximal inhibitory concentration (IC 50 ) = 10 nM) that bind active site CypD also make...
mRNA display is an effective tool to identify high-affinity macrocyclic binders for challenging protein targets. The success of selection dependent on generating highly diverse libraries with trillions peptides. While translation elongation can canonically accommodate the 61 proteinogenic triplet codons, initiation restricted native start codon AUG. Here, we investigate ability Escherichia coli ribosome initiate 31 initiator tRNA (tRNAini) anticodon mutants at their respective cognate using...
Staphylococcus aureus (S. aureus) is an opportunistic human pathogen that causes over one million deaths around the world each year. We recently identified a family of serine hydrolases termed fluorophosphonate binding (Fphs) play important roles in lipid metabolism and colonization host. Because many these enzymes are only expressed bacteria, they valuable targets for diagnostics therapeutics. Here, we developed screened highly diverse cyclic peptide libraries using mRNA display with...
Advances in genetic code reprogramming have allowed the site-specific incorporation of noncanonical functionalities into polypeptides and proteins, providing access to wide swaths chemical space via vitro translation techniques like mRNA display. Prior efforts established that machinery can tolerate amino acids with modifications both peptide backbone side chains, greatly broadening be interrogated ligand discovery efforts. However, existing methods for confirming yield new acid building...
Prion disease is a rapidly progressive neurodegenerative disorder caused by misfolding and aggregation of the prion protein (PrP), there are currently no therapeutic options. PrP ligands could theoretically antagonize formation protecting native from or targeting it for degradation, but validated small-molecule binders have been discovered to date. We deployed variety screening methods in an effort discover PrP, including
ABSTRACT Staphylococcus aureus ( S. ) is an opportunistic human pathogen that causes over one million deaths around the world each year. We recently identified a family of serine hydrolases termed fluorophosphonate binding (Fphs) play important roles in lipid metabolism and colonization host. Because many these enzymes are only expressed bacteria, they valuable targets for diagnostics therapeutics. Here we developed screened highly diverse cyclic peptide libraries using mRNA display with...
Combinatorial library screening increasingly explores chemical space beyond the Ro5 (bRo5), which is useful for investigating "undruggable" targets but suffers compromised cellular permeability and therefore bioavailability. Moreover, structure–permeation relationships bRo5 molecules are unclear partially because high-throughput permeation measurement technology encoded combinatorial libraries still nascent. Here, we present a assay that scalable to screening. A liposomal fluorogenic azide...
Encoded combinatorial library technologies have dramatically expanded the chemical space for screening but are usually only analyzed by affinity selection binding. It would be highly advantageous to reformat outputs "one-bead-one-compound" solid-phase libraries, unlocking activity-based and cellular capabilities. Here, we describe hydrogel-encapsulated magnetic beads that enable such a transformation. Bulk emulsion polymerization of polyacrylamide hydrogel shells around microbeads yielded...
ABSTRACT Prion disease is a rapidly progressive neurodegenerative disorder caused by misfolding and aggregation of the prion protein (PrP), there are currently no therapeutic options. PrP ligands could theoretically antagonize formation protecting native from or targeting it for degradation, but validated small-molecule binders have been discovered to date. We deployed variety screening methods in an effort discover PrP, including 19 F-observed saturation transfer difference (STD) nuclear...